AUTOLOGUS REGULATION

AUTOLOGOUS REGULATION

 Our biological destiny is inherited by us in every cell. Our DNA is the repository of this information in every cell. DNA is another incredible wonder of Nature. Todd Smith gives a great description (1): Six billion base pairs of DNA are packaged into 22 pairs of chromosomes, plus two sex chromosomes. Each base pair is 34 angstroms in length (.34 nanometers, or ~0.3 billionths of a meter), so six billion base pairs (all chromosomes laid out head to toe) form a chain that's two meters long. If we could hang this DNA chain from a hook, it would be slightly taller than an average human. But that's just the DNA from one cell. Each of us have around 50 trillion cells (50,000 billion). If we took the DNA from all of those cells and laid it out in a linear fashion, it could wrap around the earth 2.5 million times, or reach to the sun and back 300 times! Yet cells manage to pack all that DNA into a structure so small we can't even see it without a microscope. 

This long hard disk is twisted and braided and compressed so amazingly in the tiny nucleus of our tiny cell. Each tiny cell contains all the information to build a complete living organism or human being. Basically it carries two types information: one for autologous regulation- continuously managing itself as per inherited temporal program. This silences most of the genes and activates only those that give it its identity and characteristics. It uses multiple layers of tools and collaborations between those tools to accomplish this as per instructions embedded in it. Second type of information is its mesh like behaviour. Cell functions on its own as per the type of cell it becomes but it also functions collectively with other cells to build as per the design of the organism. On top of this mesh there are non cellular players like bioelectrical networks that influence each cell and also collectively as another language of communication amongst them. 

Cells are very crowded places: there are some 42 million protein molecules in a simple cell, revealed a team of researchers led by Grant Brown, a biochemistry professor in the University of Toronto's Donnelly Centre for Cellular and Biomolecular Research. The majority of proteins exist within a narrow range -- between 1000 and 10,000 molecules. Some are outstandingly plentiful at more than half a million copies, while others exist in fewer than 10 molecules in a cell. These molecules move very very fast inside the cell. In a blog by Ken Shiriff where he quotes from the book Molecular Biology of the Cell: You may wonder how things get around inside cells if they are so crowded. It turns out that molecules move unimaginably quickly due to thermal motion. A small molecule such as glucose is cruising around a cell at about 250 miles per hour, while a large protein molecule is moving at 20 miles per hour. Note that these are actual speeds inside the cell, not scaled-up speeds. I'm not talking about driving through a crowded Times Square at 20 miles per hour; to scale this would be more like driving through Times Square at 20 million miles per hour!

Because cells are so crowded, molecules can't get very far without colliding with something. In fact, a molecule will collide with something billions of times a second and bounce off in a different direction. Because of this, molecules are doing a random walk through the cell and diffusing all around. A small molecule can get from one side of a cell to the other in 1/5 of a second.

 As a result of all this random motion, a typical enzyme can collide with something to react with 500,000 times every second. Watching the video, you might wonder how the different pieces just happen to move to the right place. In reality, they are covering so much ground in the cell so fast that they will be in the "right place" very frequently just by chance.

 A rendition of a cross section of a cell and how crowded it is.

 In addition, a typical protein is tumbling around, a million times per second. Imagine proteins crammed together, each rotating at 60 million RPM, with molecules slamming into them billions of times a second. This is what's going on inside a cell.

In super tiny tightly packed strands of DNA heritable intelligence decides which gene (a segment of the DNA) will be read and which part of the strands will be tightly sealed to avoid being read. The ‘reading’ of the strands is by a process using enzymes and many floppy phase changing proteins as described in previous post. So many things have to come together at the right place for the gene to be read – all inside a tiny tightly packed part of a tiny cell. 

From what is not read and what is read in our DNA a cell gets it’s identity and function. Only 10%to 20%  of the coding genes are active at any given time in a cell. There is intelligence even in the spatial arrangement of each of the 200 types of cells. It’s like each type of cell is of a particular color and shape in a puzzle and Nature arranges them to form 80 different 3-D organs with incredible functions like our eyes which allow us to see and liver that does complex processing. Cells also form bones and cartilage and tendons and muscles. All of this different things made from the same basic cell. And each cell can be made to turn into any other type of cell. Unbelievably each cell has information on its ‘hard disk’ to build each and every of the organs, bones, muscles and skin. We literally start from a single cell! 

As we read in my earlier post Headwaters, this ‘reading’ or  transcription of our DNA is quite pervasive and is observed in 85% of our genome. Out of this only 2% is involved in protein coding. Rest is involved in regulating this 2% and it’s translation. The more the complex organism the bigger the ratio between coding and non coding but this tells only one part of the story. 

Even in such a crowded cell with such a huge genome Nature maximizes this space by alternatively splicing 95% of the genome. So instead 50,000 genes (coding and non coding) generating 50,000 transcripts not only 85% of the genome transcribes but 98% of this transcriptome undergoes alternative splicing! Creating uncountable isoforms. By alternative splicing we mean that same region of our genome can be ‘read’ in multiple versions. Supposing we mark a region from 1 to 10 and  neighbouring region is marked from 11 to 20 as two genes but those genes due to alternative splicing can be transcribed as 5 to 9 or 3 to 6 or 2 to 9 making 3 transcripts from the same gene. This splicing can also include neighbours so it can go 7 to 15 or 3 to 12, etc. to explain it simply. So in the preceding Headwaters post we learnt about how most of the transcription from non coding regions and some proteins create layer upon layer of regulation of the protein coding genes driving the changes that make us from an egg to an adult and after puberty it launches the process of aging. Now in this post we find that that is not all that happens in the genome and it’s housing structures like histones and  chromosomes. On top of this there is spliceosome that cuts up the genome into not just linear transcripts across its length but unending variety of isoforms due to rampant alternative splicing. Look at the packaging brilliance of Nature: a 2 meter long DNA 85% of which transcribes into transcripts in a nucleus that is 10 microns (one micron is one millionth of a meter) would be miraculous enough but Nature maximizes this by adding pervasive alternative splicing that creates multiple transcripts from same gene! Thereby multiplying the number of transcripts by multifold that are produced from the 2 meters. 

 

 

From Universal Alternative Splicing of Non Coding Exons by Tim Mercer et. al

 Only a limited number of transcripts whether of a full gene or alternatively spliced gene translates into protein. In my previous post Headwaters we read about how these shapeless, floppy proteins gather near a gene activation site and magically phase change into a condensate that hovers over the site. Similarly a different condensate activates splicingAn article published in Genome Biology Journal on 28^th November 2018 by Dr. Steven Salzburg et. Al. states the following: “We assembled the sequences from deep RNA sequencing experiments by the Genotype-Tissue Expression (GTEx) project, to create a new catalog of human genes and transcripts, called CHESS. The new database contains 42,611 genes, of which 20,352 are potentially protein-coding and 22,259 are noncoding, and a total of 323,258 transcripts. These include 224 novel protein-coding genes and 116,156 novel transcripts. We detected over 30 million additional transcripts at more than 650,000 genomic loci, nearly all of which are likely nonfunctional, revealing a heretofore unappreciated amount of transcriptional noise in human cells.”

The interesting thing to note is the huge number of transcripts they found: 30 million! They claim that most of them are non functional but Nature rarely spends resources to construct huge volumes of non-function things. The non protein coding transcripts too have very important roles. In a paper titled ‘Pervasive Transcription of the Human Genome Produces Thousands of Previously Unidentified Long Intergenic Noncoding RNAs’ by Matthew J. Hangaue et. Al. the authors say “It is now becoming more and more clear instead that, far from being genetic “deadwood” these repetitive expanses are actively and deliberately transcribed into non-coding RNAs which play a major role in regulating gene expression and silencing, organizing nuclear architecture, compartmentalizing the nucleus, and modulating protein function.” My previous post explains in detail the various types of non coding transcripts and the regulatory roles they play but here we additionally examined the alternative splicing that generates not only variety of coding transcripts but also as we read above huge number of non coding transcripts. 

 What is fascinating is how these transcripts govern their own births: if you recall we covered Long non coding RNAs in the previous post-in a paper titled ‘Epigenetic regulation of alternative splicing: How LncRNAs tailor the message’ by authors Pisignano and Lafomery they write about some of the ways in which LncRNAs regulate alternative splicing which in turn leads to various transcripts including LncRNAs. An excerpt from their paper “Both short (<200 nt) and long (>200 nt) non-coding RNAs can contribute to the regulation of alternative splicing in many different ways; either indirectly by regulating the activity of splice factors; or directly, by interacting with pre-mRNAs. Long non-coding RNAs (lncRNAs) are particularly well suited to these roles due to their demonstrated capacity to act as regulatory molecules that modulate gene expression at every level. Either alone, or in association with partner proteins, these long RNA polymerase II transcripts have been shown to take part in a wide range of developmental processes and disease in complex organisms.” So which are the ways they mentioned in which LncRNAs regulate alternative splicing:

1.     LncRNAs regulate alternative splicing through chromatin modification: An intimate relationship exists between lncRNAs and chromatin conformation.  LncRNAs regulate chromatin modifications by recruiting or directly interacting with histone-modifying complexes or enzymes at specific chromosomal loci. A possible lncRNA-mediated crosstalk between histone modifications and the pre-mRNA splicing machinery has also been proposed. Several lncRNAs appear to control important aspects of chromatin organization including chromatin looping, either remaining tethered to the site of transcription or moving over distant loci. 

2.     LncRNAs regulate pre-mRNA splicing through RNA-DNA interactions: LncRNAs can tether DNA forming an RNA-dsDNA triplex by targeting specific DNA sequences and inserting themselves as a third strand into the major groove of the DNA duplex. These are known as R-loops; three-stranded nucleic acid structures, composed of RNA–DNA hybrids, frequently formed during transcription. Aberrant R-loops are generally associated with DNA damage, transcription elongation defects, hyper-recombination and genome instability. Recent lines of evidence indicate a potential role for R-loops in alternative pre-mRNA splicing. A class of lncRNAs, the so-called circular RNAs (circRNAs) are abundant, conserved transcripts originate from a non-canonical AS process (back-splicing) leading to the formation of head-to-tail splice junctions, joined together to form circular transcripts. 

3.     LncRNAs regulate pre-mRNA splicing through RNA-RNA interactions: Identified in multiple eukaryotes, Natural Antisense Transcripts (NATs) are a class of long non-coding RNA molecules, transcribed from both coding and non-coding genes on the opposite strand of protein-coding ones. Regardless of their genomic origin, NATs can hybridize with pre-mRNAs and form RNA-RNA duplexes. In some cases, a double function is also possible, and NATs can encode for proteins on one hand, while at the same time working as non-coding molecules modulating the splicing of a neighbouring gene’s transcript. 

4.     LncRNAs regulate pre-mRNA splicing by modulating the activity of Splicing Factors: lncRNAs interact in a dynamic network with many SFs and their pre-mRNA target sequences to modulate transcriptome reprogramming in eukaryotes. LncRNAs regulate the localization and phosphorylation status of Splicing Factors. 

 The authors conclude by stating that “With the increasing prevalence of splicing events and the discovery of over a hundred thousand lncRNAs, it is likely that the involvement of lncRNAs in regulating AS is far greater than the currently known.”

 

  

Regulation of pre-mRNA splicing by lncRNAs. LncRNAs (red) are able to control pre-mRNA splicing by (a) modifying chromatin accessibility through recruiting or impeding access to chromatin modifying complexes at the transcribed genomic locus. In some cases, this might result in more drastic long-range structural changes; (b) interacting with the transcribed genomic locus through an RNA-DNA hybrid; (c) hybridizing with the pre-mRNA molecule (light blue); (d) promoting SF recruitment or by sequestering SFs into specific subnuclear compartments, thereby interfering with SF activities. Credit: Epigenetic Regulation of Alternative Splicing: How LncRNAs Tailor the Message. Authors: Giuseppina Pisignano and Michael Ladomery. 

In my preceding post Headwaters we see various ways in which many types of non coding RNAs regulate gene expression not only inside the cell but also through the circulating secretome. Here we saw how alternative splicing leads to protein diversity and non coding transcription by creating alternative transcripts from the same gene. But what is amazing is that non coding RNAs influence the alternative spliceasome. A very interesting paper titled Aging is associated with a systemic length-associated transcriptome imbalance by Dr. Luis Amaral et. Al. in which they find out that as we age longer transcripts reduce and many of them are associated with longevity genes. They cite various possible causes as the source of the origin of these change like heat shock protein leaving translation with truncated protein lengths and spliceosome and splice factors deliberately transcribing shorter transcripts. But the best clue is that they also found in some subset of tissues and cell types exact opposite is seen happening! In these short transcripts are seen reducing and long transcripts are seen increasing. So what is this a dead giveaway of? Temporal program of autologous regulation. The age related changes are not random but are orchestrated by transcription and splicing machinery and their coplayers. In a paper titled Aging associated changes in the expression of LncRNAs in human tissues reflect a transcriptional modulation of ageing pathways by Dr. Joao Pedro de Megalhaes et. Al they observed that LncRNAs are very tissue and lineage specific and typically highly specific spatio-temporal expression patterns. This again shows evidence of an intricately designed regulatory plan that unfolds with timeline of the living organisms. All this intricately complex regulation in such tiny environment is for spatial and temporal organization of a life form:

Spatial organization: Imagine a tiny cell 1/10^th the diameter of a human hair has information that it reads which tells it where it should locate itself with respect to other cells in our body. So a cell that is designated to be an eye cell, as it emerges from the multiplication of cells from one single fertilized egg, knows it has to move precisely towards the sockets being formed in the head and then through epigenetic changes it becomes an eye cell! It will not float and land up on the hand or turn into a skin cell in the eye. The precision is mind boggling. Where is that information, that instruction that it must move there to become an eye cell? It’s already labeled in its DNA. Imagine tens of trillions of cells each knowing exactly where it needs to locate itself in a 3 dimensional space of the life form and then what it needs to become to form various organs and tissues and muscles and bones! It must need to coordinate and jostle with its neighbours to land at its physical destination. Dr. Michael Levin says there is Bioelectrical memory which connects all cells in a mesh and guides each cell to where it needs to be. This process is called spatial organization. 

Temporal organization: Once a cell takes its place and it’s epigenetic buttons are clicked to transform it into a type of cell a whole different process of organization begins. In this process the 10% or 20% of the coding  genes which typically are active begin to print proteins that fulfill their various tasks in line with their cell’s type. So a pancreatic cell with code for insulin for example. These are functional tasks of the cell but parallely as we have read above there is also highly complex regulation that is happening of those protein coding genes and their transcripts. This continuous background regulation creates constant changes in the cell from birth till death. Initially these macro changes are related to development: to make us grow from an egg to an adult and after puberty the main theme of these changes is to dial down important repair and recycling systems so that within a given range the life form dies. These latter changes manifest as aging. These regulatory changes of the spliceosome, alternative splicing, non coding and coding gene transcription all together leading to a particular proteomic configuration which in turn influences the efficiency of all the tasks that are done by those proteins. The changes stop some proteins, change some proteins, reduce some proteins and increase some proteins. This is ongoing all our lives. Ironically these transcriptional and proteomic changes also affect the cells DNA itself as progressively double strand breaks increase as we age and their repair efficiency reduces when it’s needed even more. This brings us to the main observation driving this post: 

Autologus Regulation: Nature has created this unit of mind boggling complexity and intricate design: the cell. All life forms on our planet are built from this unit. Incredibly this unit produces regulators that governs itself! It produces transcripts and proteins that regulate the regulators! So basically it writes its own biological destiny. Inherited genetic factors and lifestyle factors do also influence our biological destiny but only in a narrow range. The main driver continues to remain the inherited repository of information in the cell itself. The information it carries enacts it’s spatial organization and the same source of information also enacts it’s temporal organization. It transcribes transcripts that influence the transcriptional machinery and splicing machinery to decide whether to transcribe the entire gene or whether to transcribe an alternate version or whether to silence it. Some of those transcripts along with some of the translated proteins will make further alterations to the transcriptional decisions and splicing decisions in a continuing loop of self regulation driving the two major themes: development before adulthood and aging after adulthood. Besides these two main themes there are also changes that occur due to environmental stimuli. But overall unless they are extreme or fatal these are dominated by the two main themes. Some of these instructions are exchanged between cells through direct connections with neighboring cells or through the secretions of one cell entering another. 

This self regulation is a very interesting process created by Nature which we rarely get to witness anywhere else. It’s easy to miss how incredibly remarkable is this technology developed by Nature and evolution. DNA carries information that when read sequentially builds us into an adult starting from a single cell and DNA also carries information that when read sequentially after puberty leads to gradual aging and death. We inherit both, our youth code and our death code,  from the moment we are a fertilized  egg. Let me try to explain it with an example. Let’s say a branch office is opened (cell) in which there is no manager but only an SOP manual – a standard operating procedure master handbook for the entire year that all the staff has to follow. It gives instructions to the HR dept on what kind of staff to hire. It has various printers that print out instructions daily giving tasks to all the staff. But imagine that only 30% of the employees actually do the tasks that produce the parts that the branch manufactures. 70% of the employees are getting instructions daily from the SOP to manage those 30% employees and what they produce by making changes in the master SOP that is daily giving instructions to those 30%. So the SOP itself has instructions to daily make changes in the SOP and thereby resulting in changes in the production. But those changes and their edits are so complex that it requires 70% of the employees just taking new instructions daily from the SOP and coming over and editing the future chapters of the SOP manual. These self edit instructions flow out sequentially as each new page of the SOP is read each new day of the year. Other branches also exchange data (secretome) and send their employees to also make edits in each other’s SOPs’.  In the beginning there is tremendous excitement and new teams are hired and production is going full swing making wonderful products that sell very well (puberty). At its peak the training reaches a point where a team of employees can go and open another branch (reproduction). But once that is done the SOP begins to give out instructions to edit itself (autologous regulation) so that in forthcoming pages the production quality, hiring quality, raw material quality all of it is purposely, gradually brought down (aging). In the beginning it’s hard to notice but after some months of such gradual changes the consequences begin to show and unsold products start piling up. Cash flow is affected, salaries are affected. And what at its peak was a dynamic factory full of enthusiastic, productive workers becomes demoralized and stressed out leading to even further degradation at the branch creating a snowballing stranglehold from which the branch can’t escape and at some point it shuts down which is death. This is done so that there is no over crowding of the branches creating over supply which would destroy the company itself and also to ensure fresh young staff is recruited with every new branch which is enthusiastic and hard working. 

 Coming back to our biology there are two basic goals of autologous regulation: One is to build an adult from a fertilized egg. Second is to gradually make the adult age that would culminate with sufficient degradation to cause death anywhere between average lifespan to maximum lifespan of that species. One of the key reasons for this regular recycling every generation is because thanks to a paper last year by Dr. Vadim Gladyshev we learnt of this marvelous event occurring during early embryogenesis: all the inherited errors and insults of germline cells is wiped clean to make a brand new error free baby. Have humans outgrown this need to regular recycling? Can our intelligence help us to resolve the challenge of accumulation of biological errors and insults? As mentioned in my previous post I continue to take inspiration from certain life forms which seem to be immortal in permanent youth. I cite the Ginkgo Biloba tree because a researcher Dr. Richard Dixon has studied it. Even after a thousand years the tree that he studied still had photosynthesis efficiency and immune resilience of a 20 year old tree. Question arises as to how it’s able to do this. In almost all other life forms the DNA harbors temporal instructions that, as we read above,  make changes to the spliceosome and the splicing factors and the transcription factors and the epigenetic marks which result in gradual collapse of our repair and recycling systems and ultimately death. How is Ginkgo Biloba allowing all the changes related to development to reach adulthood but freezing or blocking or erasing further regulatory changes thereby permanently remaining in youth? Many scientists wonder if we can prolong our youth would we still die when we reach 122 or 125? Ginkgo Biloba tree says no. 

Two technologies are moving towards reversing human biological age. One of them is partial reprogramming of the cell using some of the yamanaka factors. This will in effect reverse the epigenetic signature, the gene expression and the proteome back to an earlier point closer to our youth. Question is does it also change the transcriptome? If not,  aging related changes would again bring the cell back to an impaired state. If it does also turn back the temporal needle of the transcription program to where it was in our twenties then it would again take decades for the cell to get impaired again. The only catch is that this is the same path that cell would take if it were reverting back to embryogenic cell state and that state can lead to cancer. So does partial reprogramming fully protect against cancer? One can never know till many years later. Second technology is an arbitrage. Signaling and regulatory molecules circulating in the plasma of the young are injected into the circulatory system of the old. As those molecules enter the impaired cells they reset the proteome of that cell back to how it was in youth. Thereby rejuvenating those cells. This does not stop the legacy transcription in the cell which after a point would begin the degradation all over again. The question here is if the pro youth molecules are injected repeatedly would that at some point ‘flip’ the transcriptome to how it was during youth? If yes then it would take decades before the cell would get impaired again. 

Human biology is incredibly complex. But does it have to be this complex? The complexity arises to maintain autologous management of the entire body. But can it be improved? Why do we need to generate voltage only from the food we eat? Why can’t we re-engineer so that we need only sunlight for energy like trees and plants do so beautifully? So much of our body’s parts are devoted to eating, digestion and excretion. If we did not need to eat to generate electrical energy we could reduce 50% of of our organs. Also why can’t we store electrical  energy in our body/cell? We humans have created batteries to store electrical power so are we now ahead of evolution? Can we create alternate source of electrical energy in our cells? We have the intelligence to do it. Can we obviate the need for oxygen? We will also be able to edit the embryogenic process safely to alter our human organs and systems and form.  I guess all of this is possible in the distant future. It will all start with our control over biological age. 

CANCER: EXCITING CASE STUDIES OF REPURPOSED DRUG: FENBENDAZOLE

 CANCER: EXCITING CASE STUDIES OF REPURPOSED DRUG:

FENBENDAZOLE 

Please consult your oncologist and show him/her the success stories as published by Stanford oncologist before considering anything mentioned in this post. It is meant only for information purposes. 

In my earlier post I have compiled a list of repurposed drugs found effective against cancer. The beauty about this approach is that in almost all cases one does not need to change their first line therapy prescribed by their oncologist. The repurposed drug/s can be taken as an adjuvant therapy. Cancer is the most cunning and ferocious living thing on this planet. One can not expect to beat it with just a mono therapy. Repurposing drugs opens up an opportunity to safely attack it from multiple fronts ensuring much better chances of surviving it. 

This post is about a paper that came out recently in Clinical Oncology Journal by Dr. Sandy Srinivas and team from Stanford University Medical Center. This paper shares with us incredible results of using Fenbendazole on advanced stage cancer patients making them completely cancer free! Even though number of cases shared are only 3 what is astounding is that there was complete response in 3 out of 3 lasting 9 and 10 months at the time of paper. Every cancer patient should get excited by this as Fenbendazole should have a wide application across many cancers. 

Fenbendazole is a cheap, easily available anti parasitic drug used in veterinary care. In the case studies they had administered 1 gram every alternate day or 3 times a week. This dose was very well tolerated. 

                           Dr. Sandy Srinivas 

The first patient has kindly shared his name and experience with the public. Joseph Clark in his in words:

“ I had stage 4 kidney cancer that had spread to my IVC, right atrium of my heart, both lungs, pancreas, hip and spine. In April of 2019 I was deemed terminally ill with six months to live. Last ditch treatment was immunotherapy. I was given 3 half doses and it was determined that I became highly toxic from the treatment which caused pancolitis. Treatment was terminated. Now first week of August started taking Fenbendozol NO vitamins no CBD. Now second week of October 2019 MRI scans at Stanford shows my largest tumor in my left kidney was GONE! All other tumors shrinking considerably! Now January 2020 Stanford MRI scans show no evidence of disease!! (NED) all cancer gone! I’m still taking Fenbendozol 3 days on four days off consistently. STANFORD now producing Fenbendozol in there lab for human study ! I’m living proof of Fenbendozol treatment with no vitamins. It doesn’t mean vitamins won’t help but they aren’t needed to kill cancer cells either. All my records are proof positive and can be reviewed.”

 

Many repurposed drugs do show some response in cancers. I have myself listed 25 leading candidates that have shown some efficacy. But the challenge with all so far is that not yet tried in human cancer patients, showing only partial response which is transient, drug resistance sets in or toxicity in healthy cells becomes an issue for long term use. Fenbendazole seems to have cleared all these hurdles. So what makes it so successful against cancer when a thousand other attempts have failed?:

1. Cancer is quite wily and can easily outsmart a monotherapy. So for any drug to have a chance to succeed it must have a multi-modal action against cancer. Fenbendazole attacks cancer in 4 known ways:

2. It partially destabilizes microtubule polymerization. What it means is that it disrupts an important process in cancer survival which leads for some of the cancer cells towards mitotic arrest and apoptosis – means makes some of the cancer cells die.

3. It inhibits cancer cell's proteasomal machinery. This means that the garbage disposal system to remove excess proteins from cancer cells becomes disabled. This leads to build up of proteins in cancer cells leading to its death.

4. P53 is an important tumor suppressor gene. It’s protein plays a very important role in controlling cancers. For example when DNA is damaged P53 decides whether it can be repaired or whether it should go for apoptosis. If it decides it can be repaired it activates DNA repair genes to complete the task. If decision is that it’s irreparable then it activates cell self destruction. Specifically regarding cancers it supervises cell division to ensure there is no uncontrolled or too fast cell division by self destruction of such cells. In most cancers P53 is hijacked and mutated by cancer to not only stop its vital tumor suppressor activity but also convert it to help oncogenesis. So cancers convert a lethal enemy into a slave. Fenbendazole downregulates negative regulators of P53 and promotes stable transcriptional activity – basically reactivating the hijacked and redundant P53 gene and its production of tumor suppressor proteins.

5. Fenbendazole also downregulates glycolitic enzymes crucial for cancer cell survival. Basically because cancer cells undergo feverish activity they need a lot of glucose for powering these furious actions. If this supply of glucose is disrupted in anyway cancer cells would die due to all loss of energy. 

6. Many drugs do succeed but the cancer cell causes overexpression of MDR-1 gene leading to more production of p-glycoprotein. This results in drug resistance so all the benefit seen initially with a drug against cancer begins to fade away. In the case of Fenbendozole it is not a Pgp substrate so does not succumb to drug resistance. 

7. Other drugs are efficacious against some cancers, avoid drug resistance too but turn out to be toxic to healthy cells beyond certain doses. This limits their use. Fenbendazole is not toxic and very well tolerated at the dose required for eliminating cancers. 

The above combination of fortuitous factors enables Fenbendazole to show such miraculous success in completely eliminating cancers. So why isn’t this made available to all cancer patients as a prescription drug? Well as it can not be patented no big Pharma has any interest in spending huge amounts of money getting such FDA approval. That’s why this paper from Stanford is valuable as it gives credibility to Fenbendazole and allows some doctors to consider it as an adjuvant therapy for terminally ill cancer patients.

So how did it all start for Fenbendazole against cancer and just who is Joe Tippens?:

Joe Tippens has a successful career in finance and private equity so one can count him as a credible source. He was diagnosed with small cell lung cancer in 2016. He was being treated at MD Anderson hospital but his cancer had metastasized and spread to his neck, right lung, stomach, liver, bladder, pancreas and tail bone. Joe was told in 2017 that he had only 3 months to live. In all such stories there is always some instance that makes you feel was UT destiny? He noticed a post on a sports board simply saying if you have cancer contact me. Now the person who posted it, a veterinarian, was known to Joe from many years so he immediately called him. He told Joe that a scientist at Merck Animal Health had performed cancer research on mice and stumbled upon a veterinarian drug that was destroying various cancers fully. That Merck scientist herself got diagnosed with stage 4 brain cancer was also told like Joe that you have no hope and 3 months to live. The scientist started taking the veterinary medicine called Panacur-C which had in its 1 gram, 222gms of Fenbendozole as the active ingredient. Six weeks later she was cancer free. So hearing this story Joe began to take it for 3 days with 4 days holiday then take it again for three days and so on along with bioavailable curcumin and CBD oil taken every day from 3rd week of January 2017. He had no side effects at all and now believes that Fenbendazole of similar dose can be taken everyday without a break of 4 days. When he took a PET scan on May 2017 at the hospital his oncologist was shocked to see an all clear scan! In Joe’s own words:

 

                             Joe Tippens

"Are you kidding me? 3 months earlier, In January, my PET lit up like a Christmas tree.  There was cancer in my body from head to toe.  And it was a terrifyingly dangerous metastasis that leaves virtually 100% of its victims dead within 3 months.  Here I was 3 months later and the PET scan was completely dark......void of any light.....anywhere.”

Joe continues to take Fenbendozole and continues to remain cancer free till date. When created a blog https://www.mycancerstory.rocks/home/categories/my-story it created a global storm with Panacur-C running out of stock. His story is what also inspired the 3 patients in the Stanford paper mentioned above to take Fenbendazole.  While he has compiled around 50 success stories with Fenbendazole other than him he does mention that he has come across a few that did not show much response with it. So it is not a total cancer cure. But for every late stage cancer patient, with its safety profile, question is why should they not try it?

HEADWATERS: FROM WHERE THE AGING CASCADE BEGINS

ETERNAL YOUTH 

HEADWATERS

From Where The Aging Cascade Begins

One of the greatest mysteries of biology is the source of sequential orchestration of genetic events in the DNA. For example when we are around 6 years old we begin to lose our milk teeth. What tells exactly at that age to do this? What decides the timing for puberty or menopause? Or just after puberty what tells our cells around that age to bring down the protein production support machinery's efficiency by 70% to begin aging? As stated in my earlier posts there are deliberate very harmful changes that occur in a timed manner to promote aging. If we can find out from where all these instructions come may be the detrimental ones could become a therapeutic or gene therapy target. Is it a single part of our brain that gives these instructions at a particular time/age? No. Apparently each of our 30 trillion cells has its own manager that releases time regulated instructions or temporal regulation. So it seems there is no single boss but 30 trillion managers each managing their own cell and coordinating with other managers. There is a postal system enabled by our blood circulation that scurries messages amongst all the managers probably to react to environmental stimuli or to even out the changes in gene expression. The messages in this postal system are carried in envelopes: EVs: extracellular vesicles and their cargo are the messages. In our body something keeps time and releases instructions for developmental changes in the first phase and aging related in the next phase. No one has been able to point out from where. Even the scientists who are computing clocks that measure changes in methylation in our genome, epigenome and rDNA also have not been able pin point the source of these changes. We will try to do some detective work but later in this essay, next we will share some of the mindblowing discoveries and their movies being made showing us for the first time how genes are turned on. It is important for our quest to understand this.

The brilliant scientist Ibrahim Cisse. Bryce Vickmark

Ibrahim Cisse is a great success story. From a background with few resources in Niger, Africa through education and merit he is today a biophysicist at MIT, USA. His tweak to single cell high resolution microscopy allowed him to take films of RNA transcription in action. This has changed the way we thought about how genes were activated. It seems there are lots of tiny floppy proteins that coalesce into phase change droplets at a target gene switch. They form a mesh in which many proteins would flit in and out sometimes within seconds. They would collectively turn on a gene. The length of their stay would determine how many proteins would get transcribed. Once the job is done these floppy proteins would disperse like a flash mob.

It takes a village of proteins to turn on genes. W.K. Cho Science 2018

It is indeed fascinating to see how the required proteins gather at the required address. Then on their own like magic condense into droplets that coalesce together to form a new village of fast moving droplets. In this temporary body other proteins flit in do their job and zip out sometimes within seconds. Where do these transcription proteins come from? What gives each of those proteins the instructions to all of these complex tasks which would seem to need some level of intelligence at such nano scale?! And voila the selected gene is turned on and begins transcribing a copy of itself. These required proteins do not have a Google Map (t) that home them in at the right address in the looong DNA coils. They bump around like manic blind mice till they bind into the lock made for them. It’s so tempting to believe that some central intelligence is guiding and sequencing these trillions of gene controls of activation or silencing or methylation or acetylation, etc. But such a complex central control tower would be unmanageable. So this becomes our clue as to source of all instructions. We will come back to it later. Next let us review our DNA. The proteins as one saw execute almost all our processes. There may be 90,000 types of proteins. Each one being born in their designated cells and doing their given tasks. Where do proteins come from? Our DNA but only 3.5% of our DNA is protein coding. Until recently scientists used to consider the balance 96.5% as junk DNA. But DNA formation is the culmination of hundreds of millions of years of adaptations and optimizations. There is no way so much DNA would be wasted from generation to generation. Recent discoveries have shown that the non coding 96.5% carries out some of the most important functions. Our body is a collection of cells. Each cell is same and born with identical DNA ‘brain’. Right from the time they differentiate into the 200 different type of cells giving our body distinct organs or structure. Till they die they follow instructions. The instructions come either from within its coding DNA or non coding DNA or from other cells. So just as I formed a conclusion about how aging is executed in our body I have also come to believe that the answer to the greatest mystery of biology is that which instructions will reach which cell and when is decided by the cells themselves as per Nature’s program carried in the non coding section of the DNA. Now I want you to imagine that there are 30 trillion offices around the world. Each office has a central computer with pre-installed software and a 3D printer (transcription/translation). Each office gets designated to a department (organs, teeth, bone, etc.) All the offices are connected by internet. Internet is the signalling system. This is a true democracy so no concept of Leaders. Each office is it’s own leader and co-exists and coordinates with other leaders. Now all the offices have a common instruction manual but the pre installed software controls which pages are visible/actionable and which pages can not be opened. The pre-installed software in each office activates tasks which result in the smooth operation of the entire company. Each one does it’s part and collectively the software orchestrates the functioning of a complex working enterprise. Our DNA is the preinstalled software in each cell (office). Only 3.5% of the software can assign printing jobs that result in posting of instructions/actionable items. Rest of the software is about making sure by individually executing orders it collectively runs our entire body and all its systems and functions. It’s like a live jigsaw puzzle of 30 trillion parts and each part has a chip that ensures it goes and locks on its own at its correct location in the puzzle. If all cells carry the same blue print how do cells differentiate into the 200 different types and how do they function in the role assigned and not some other role? It is done by control of gene expression. At any given time only a few genes are active in a cell approximately 3% to 5%. So how is cellular type, it’s function and it’s production of proteins which in turn control various processes in our body controlled or regulated? There are various mechanisms of gene regulation, structural as well as chemical. Like Chromatin accessibility, histone modifications including acetylation and deacetylation, ubiquitination, phosphorylation, etc., DNA methylation, demethylation, binding affinity alterations, repressors, during transcription, during transport of mRNA, stability of mRNA, during translation and postranslational, etc. As one can see the regulation of genes is highly complex but works in concert with regulation of genes in other cells to culminate into the object of such activity both in the cell and collectively in the body. We are built and we operate based on these controls. Overall there are  three basic types of changes that occur due to regulations of gene expression: developmental changes which build us from a single egg to an adult, stimuli adaptative changes and finally aging related changes (post developmental changes). As soon as developmental changes stop just a little after puberty there are deliberate negative or harmful changes that begin our multi decade process of aging. These changes are also highly conserved to ensure that all humans degrade to a point of death. But the question still remains: From where do the instructions come to execute exactly the control action (these changes) needed in each cell at exactly the time it is needed?

Now we have seen above what a fascinating process it is of activating a gene. The activation and repression of genes in each cells DNA culminates in our creation and operation. What scientists have not yet been able identify so far is from where do activation repression instructions come. Not only that the non stimulus related instructions come in a sequential manner. If it didn’t we would turn 20 then suddenly turn old then turn back into 7 year old, randomly. Another aspect of the mystery is how correct set of genes are switched on and off at exactly the correct location. If not eyes would appear on chest and finger could grow on skull. What is incredible is the low error rate over 200+ million years. The fidelity and precision of the system is mind boggling. As mentioned above controlling millions of complex processes in trillions of cells is impossible to achieve from a single source like a part of our brain. Its would be 100000 times more complex than managing all the flights in the world from a single air control tower. So we can infer that the origin of the instructions are also decentralized. In that case the only place in a cell that contains such data is our DNA. Now we have researched quite a bit on the coding part of our DNA. From that we can conclude that coding part needs upstream instructions to execute it’s control over gene expression and protein creation. From the little we have studied the 96.5% non coding part of the DNA we have noticed some interesting functionalities. The majority of the non coding part of the DNA (which does not print proteins) is highly conserved over 200 million + years. From the research papers that I read it hinted at this portion of DNA controlling the part of ‘which’ instruction will be executed ‘where’ and most importantly ‘When’. The when part ensures we move from a baby to an adolescent and later an elderly person in a sequential manner not suddenly becoming old then turning baby then turning middle age or any such random order. This clock that decides when to trigger which change and where lies hidden and protected in depths of our non coding DNA. There was an ongoing debate wherein one side said that almost all the non coding part of DNA is junk accumulated over the years and has no function. Edward Rubin's team at Lawrence Berkeley National Laboratory snipped out 3% of the non coding DNA in mice and did not find any abnormalities. On the other side Martin Sauvageau and colleagues at Harvard University and Broad Institute found that when they created a knockout mice model without 18 Long Non Coding RNAs (LncRNAs) it caused major growth defects including abnormalities in lungs, heart, gastrointestinal tract and neocortex. While a deletion in a small part of non coding DNA not showing any abnormalities does not prove that rest of the non coding DNA has no function. Whereas deleting a part of non coding RNAs causing fatal abnormalities does prove that non coding sections of DNA have critical functions. Hundreds of studies recently have uncovered more and more functions of the non coding elements. Data from ENCODE suggests that more than 75% of the human genome is transcribed into RNAs, whereas only 3% of these RNAs are from protein coding genes (Djebali et al., 2012; Ecker, 2012; Pennisi, 2012). The balance 72% transcribed RNAs will have functions most of which are yet to be discovered.

This is how we visualize DNA

But this is how it functions with layers of regulation

There are about 20,000 genes in the human genome, but as many as 1 million of regulatory elements in the non coding part of DNA.  If we compare the number of protein-encoding genes in worm and human, for example, humans don’t have that many more protein-coding genes than worms. The noncoding genome scales up much better with the developmental and pathological complexity of an organism. The fraction of protein-coding DNA in the genome decreases with increasing organismal complexity. In bacteria, about 90% of the genome codes for proteins. This number drops off to 68% in yeast, to 23-24% in nematodes and to 1.5-2% (or 3.5% as per some studies) in mammals. Using data from comparative genetic studies, the researchers found that the 300,000 functional elements they found made up about 70 per cent of the evolutionarily conserved non-coding DNA shared by mice and humans. The spatial organization is how regulatory elements know where to execute their tasks. How the regulatory elements contribute to activate a gene is not determined by a specific recognition tag, but by where precisely the gene is in the genome says scientist Francois Spitz. The winding and folding of the DNA around histones and nucleosomes which fold again to form a 30nm fiber which forms loops called Chromatin which again regulates transcription by remaining tightly condensed or open for allowing the village of phase change proteins to activate genes. All this folding is not stochastic but precise. The control of regulation occurs due to specific addresses/locations. Any misfolding can result in incorrect genetic actions. The non coding DNA controls our biological cycle by structural and chemical manipulations. The million non coding regulatory elements are interspersed with coding regulatory elements there are co-regulators and there are regulatory elements that control other regulatory elements which sometimes in turn control other regulatory elements. There are hundreds of different mechanisms of manipulating transcriptional activity acting individually or in clusters. Creating a complex web of intricately managed cellular and biological regulation. This complex management leads us to grow from an egg into an adult and from young to old. The non coding part of our DNA starts this complex regulatory cascade.
Some of the important elements of non coding DNA – this is not an exhaustive listing but to show the massive amount of regulation that originates from the non coding part of DNA and also to see how complex is its control over our biological life:

Transposons: I would like to quote from a very good essay by Francesca Tomasi and Olivia Rhoades: Nearly 46% of our DNA is made up of transposons! For millions of years, transposons enjoyed plenty of travel around our genome. They inserted themselves throughout our evolving DNA for as long as they could before these changes started to make the host human less suited for survival in a given environment. When their random insertion provided some sort of life advantage—increased ability to absorb certain nutrients, for instance—or took place with no negative effect, the resulting modifications to the genome were passed on to future generations. Any insertions that caused death or illness, meanwhile, were a lot less likely to make it past a single generation. As such, over millions and millions of years of trial and error, transposons gradually integrated themselves in increasing numbers throughout our genomes. Eventually, their ability to move without negative consequence likely became, for the most part, saturated. And as a result, over 99% of the transposons in the human genome lost their ability to move. But we still have some active transposable elements within us: sometimes they can wreak havoc and cause disease. At a much finer level of resolution, transposons contribute to creating genes, modifying them, and programming and reprogramming them. Many transposons and retroelements contain captured gene fragments and can be part of gene regulatory regions. The bottom line for genomes is that the cleavage and resection of DNA by transposases virtually guarantees sequence variation, genome scrambling, and the appearance of transposons at rearrangement breakpoints. Simply put, transposases drive genome evolution.

Non Coding RNA: a good reference is a chapter: Loudu Srijyothi, Saravanaraman Ponne, Talukdar Prathama, Cheemala Ashok and Sudhakar Baluchamy (October 10th 2018). Roles of Non-Coding RNAs in Transcriptional Regulation, Transcriptional and Post-transcriptional Regulation, Kais Ghedira, IntechOpen, DOI: 10.5772/intechopen.76125. Available from: https://www.intechopen.com/books/transcriptional-and-post-transcriptional-regulation/roles-of-non-coding-rnas-in-transcriptional-regulation.
Non coding RNAs are functional RNA molecules from our non coding DNA but do not code proteins. There are many types of non coding RNA: such as small non coding RNAs – sncRNAs: miRNA, piRNA, SiRNA, SnRNA and long non coding RNAs – lncRNA: lincRNA, NAT, eRNA, circRNA, ceRNA, PROMPTS. ncRNAs play critical roles in defining DNA methylation patterns as well as chromatin remodeling this having a substantial effect on epigenetic signaling. ncRNAs play roles in transcriptional and post transcriptional regulation. Methylation patterns change in a linear fashion through out our life and to an extent where they are used by algorithms to predict biological age. ncRNA regulation is tissue specific and also makes changes in a linear fashion following our stages of lifecycle: earlier ensuring developmental changes and just after puberty age related changes.
Find below a diagram encompassing the RNA universe:

Small interfering RNA – siRNA and micro RNA- miRNA: These molecules induce mRNA degradation or translational repression which thereby changes gene expression. Surprisingly about 60% of the translated protein coding genes are negatively regulated by miRNAs! To make it even more complex there are lncRNAs which bind and degrade target miRNAs thereby upregulating that gene's expression. Layer upon layer of regulation. miRNAs also play role in cell proliferation, cell differentiation, development and cell death.
Long non coding RNAs: Their actions can be divided into 4 types. The diagram below will explain them:

LncRNAs have diverse regulatory functions and might regulate gene expression by modulating chromatin remodeling, cis and trans gene expression, gene transcription, post-transcriptional regulation, translation, protein trafficking and cellular signaling. These below are some of the ways in which lncRNAs regulate:
Transcriptional regulation is done by:
Enhancer ncRNAs – eRNAs – as name suggests they upregulate gene expression.
Activating ncRNAs – transcriptional activating function. Although function is similar to eRNAs their mechanisms are different.
lncRNAs that recruit chromatin modifiers- they recruit chromatin remodeling complexes to specific DNA location to activate or repress genes.
ncRNAs involved in genomic imprinting- participate in epigenetic silencing of an allele inherited from either parent. One example is X-chromosome inactivation in females.
Post translational regulation is done by acting as competing endogenous RNAs that regulate microRNA levels which in turn modulate mRNA levels by altering mRNA stability, mRNA decay, and translation. Some of their regulatory actions:
LncRNAs as a source of miRNAs- 50% of the miRNAs are produced from non coding transcripts. LncRNA genes contain embedded miRNA sequences which may be located within an exon or an intron or occur in clusters within the genome. Though the sources are different, the pathways converge at the level of pre-miRNA structure which produce miRNA.
LncRNAs as negative regulator of miRNA- as mentioned above miRNAs act as negative regulator of gene expression lncRNAs competitively bind them and degrade them thereby upregulating target gene expression.
LncRNA mediated mRNA degradation- they do this mRNA degradation directly independent of miRNAs.

Cis Regulatory Elements: Non Coding regulatory elements near a gene. Cis-regulatory events are complex processes that involve chromatin accessibility, transcription factor binding, DNA methylation, histone modifications, and the interactions between them, control of chromosomal replication,  condensation, pairing and segregation. Types:
Promoter: helps in Initiating transcription of a gene.
Enhancers: provide binding site to Transcription Factors to enhance gene expression.
Silencers: repress the gene after binding with Transcription Factors.
Response Elements: provide locational homing for Transcription Factors.
Insulators: Acts as a boundary wall.
Almost 1/3rd of the genome- about 1 billion base pairs – may be involved in cis-regulatory functions.

Trans Regulatory Elements: modify expression of distant genes.

Introns: Introns are non coding elements inside a gene which are spliced out before transcription of the remaining gene (exon). Alternative splicing allows multiple proteins to be generated from the same gene. 90% of our protein coding genes have introns and of those 95% have alternative splicing! Human genome contains an average of 8.4 introns/gene: 139,480 in our entire genome. Accounts for 25% of our genome. Why our genome has so many conserved Introns is still being discovered but some of the regulatory functions that have been found are transcription initiation, transcription termination, time delay during transcription, alternative splicing, recruitment of nuclear export factors, recruitment of shuttle proteins, it increases translation yields, etc.

Repeated non coding DNA sequences: repeated noncoding DNA sequences at the ends of chromosomes form telomeres. Telomeres protect the ends of chromosomes from being degraded during the copying of genetic material. Repetitive noncoding DNA sequences also form satellite DNA, which is a part of other structural elements. Satellite DNA is the basis of the centromere, which is the constriction point of the X-shaped chromosome pair. Satellite DNA also forms heterochromatin, which is densely packed DNA that is important for controlling gene activity and maintaining the structure of chromosomes.

As ENCODE data suggests that 75% of the human genome is transcribed to RNAs but only 3% of that is from protein coding genes. So rest of the RNA and other factors have complex functions as mentioned above – most of which are yet to be discovered. We read above how multiple transcription factors help activate a gene. Many of these transcription factors are coded in the non protein coding part of our DNA. Each has a function creating an affinity for co-factors, their shapes locking to the exact location. There are regulatory factors that regulate other regulatory factors. So for example one example of factors would regulate as per their purpose but then another layer of factors emerge due to temporal reasons as during aging to bind and block the first layer from activating genes. Such factors emerging from transcription of non coding DNA seem to regulate at various levels, pre-transcription, post transcription, post translation, etc. Both spatial and temporal organization of genome is incredible but what fascinates me is the temporal organization of genomic activity. All the little information we have discovered about the world of non coding DNA shows incredibly complex and precise management of our body through the management of the intracellular and extracellular environment through the management of gene expression and post gene expression. To go one step ahead one can say that which proteins get printed correctly, are stable and get activated and which proteins are blocked at any given time in our lifecycle determines the homeostatic status of our various systems which culminates into our lifestage at that time. The non coding elements not only regulate gene expression/repression but also ultimately protein production/repression.  Our biological destiny is defined by the proteome which is regulated by the transcriptome. The 90,000 types of proteins that are produced, which help run our bodies, from the 3.5% of the genome are regulated by the factors transcribed from 96.5% of the genome! We can safely deduce that the highly conserved part of the non coding genome holds the ‘clock’ that triggers various transcriptions based on time or life stage. This temporal execution turns us from an egg to an adult and then gradually and deliberately in a calibrated manner destabilizes homeostatic balance and efficiency of important processes like repair to make us grow old and eventually die. Just like the developmental program, the aging program too is global, unfolding in every cell in our body. Change is constant in our lifecycle. I am not talking about the changes that occur for day to day operations. I am talking about macro global changes. If there were no changes we would remain an egg. We would remain a baby if changes stopped after we developed into one. These changes make us an adult and then begin aging till we die. Now here is what is fascinating: If we stop these lifecycle changes from occurring after we become an adult there is no reason why we could not remain young forever. This is not a theoretical speculation. Researcher Richard Dixon from University of North Texas and collaborators discovered that a 1,000 year old Gingko Biloba tree's gene expression was the same as a 20 year old tree with no sign of senescence or deterioration. The tree's ability to photosynthesize, germinate seeds, grow leaves or resist disease/infections remains the same as trees thousand years younger.

1,400 year old Gingko Biloba tree planted by Chinese Emperor of Tang Dynasty (618-907) in full bloom at Zen Monastery in Shaanxi!

Our biological systems are so brilliantly designed that at its homeostatic peak it could maintain optimum operations almost forever. If the Gingko Biloba tree can figure it out why can't we. If we want to remain forever in homeostatic bliss and at our youthful peak we will need to discover the region of non coding DNA that gives birth to elements that trigger the various changes that lead to the aging phenotype. Once discovered we would need to either edit or block those regions or elements. We would need to freeze our gene expression pattern as soon as we become an adult. Reward will be eternal youth. This, whenever does happen, would be a permanent solution to remaining young but what about now? Another strategy is to hack the factors and proteins that cause progressive age related changes in the activation and repression of genes and replace them with factors and proteins from a young environment. This will change the gene expression signature back to what it was in youth. In turn that would make us young again. Only catch is that unlike the permanent change to youth that is possible by discovering the birthplace of elements that make the age related changes in gene expression, the hacking protocol requires regular hacks for life to maintain youth.

ATTACK CANCER TODAY BY APPROVED DRUG REPURPOSING

ATTACK CANCER TODAY BY APPROVED DRUG REPURPOSING

There are many sites that report cancer studies. Unfortunately cancer drugs take years before they can move from lab to prescriptions - as many as 10 years and only 10% or less can get FDA approval. Cancer mutates at manic pace and cancer patients do not have time. Through this blog I try to bring therapeutics that the patient can get administered today. Earlier post comprehensively covers natural molecules and compounds which have shown success in studies conducted by reputable labs against cancer. This blog was also amongst the very early ones to predict the importance of cancer immunotherapies.

In this post I am listing various approved therapeutics that a cancer patient can ask their doctor today to prescribe to them to combat their cancer. Recent major discoveries have shown drugs approved by FDA for a particular disease proving highly beneficial in another disease. Two greatest examples of such drug repurposing are Sidenafil originally approved for heart conditions and now prescribed for erectile dysfunction. The second example is Metformin prescribed for type 2 diabetes and recently found to have healthspan and lifespan extension abilities. Drug repurposing can be a very powerful weapon in the hands of cancer patients as it opens up hundreds of new relatively safe options that they can use right away to fight cancer.

Below are listed some such candidates:

1. Drug Cocktail: Wan L. et al. showed that a combination of 3 FDA-approved drugs, the over-the-counter drug aspirin, the well-known antibiotic doxycycline and mifepristone (a progesterone receptor antagonist known as an abortifacient pill) used together with the amino acid lysine could effectively and safely prevent cancer metastasis. Now metastasis or spread of cancer from its original site causes 90% of the cancer related deaths so this discovery is very valuable for cancer patients who are in early stages or can also mitigate recurrence. They stopped adhesion of cancer cell lines to either endothelial cells or extracellular matrix via down-regulating cell adhesion molecules ICAM-1 and α4-integrin. Without cancer cells being able to stick to blood vessel cells they can not rupture them and spread. In their in vivo experiment, a four-day pre-treatment followed by a 30-day oral administration of the quadruplet drug combination to mice inoculated with melanoma cells produced significant inhibition of cancer metastasis in the lung dose-dependently without any marked side effects. But all 4 need to be taken together for their synergistic benefit.

2. Nitazoxanide: an FDA-approved antiprotozoal drug with excellent pharmacokinetic and safety profile, is the only molecule among the screening hits that reaches high plasma concentrations persisting for up to a few hours after single oral dose. Nitazoxanide activated the AMPK pathway and downregulated c-Myc, mTOR, and Wnt signaling at clinically achievable concentrations. Nitazoxanide is given in combination with either irinotecan and Ketoconazole. Senkowski et al had in vivo success by combining it with irinotecan. Nazir M. developed a screening that identified the antifungal agent ketoconazole  as  selectively  toxic  to  hypoxic  and  nutrient  deprived  cancer  cells  when  combined with  nitazoxanide.

3. Itraconazole: is a broad-spectrum anti-fungal agent. An emerging body of in vivo, in vitro and clinical evidence have confirmed that it also possesses antineoplastic activities and has a synergistic action when combined with other chemotherapeutic agents. It acts via several mechanisms to prevent tumour growth, including inhibition of the Hedgehog pathway, prevention of angiogenesis, decreased endothelial cell proliferation, cell cycle arrest and induction of auto-phagocytosis. These allow itraconazole, either alone or in combination with other cytotoxic agents, to increase drug efficacy and overcome drug resistance. 
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5588108/#!po=27.9762

This above is a link to review article by Rachel Pounds et al. It gives response data of patients with the following cancers: ovarian, prostrate, breast, lung, bcc, pancreatic, biliary tract, mycosis fungoides and acute leukaemia.

4. Mebendazole: What do an anti-parasitic drug used to treat pinworms and a frequently used anti-malarial drug have in common? According to recent studies from the labs of Johns Hopkins and University of Kentucky investigators, both mebendazole and chloroquine could be promising medicines to combat cancer. The Brain Cancer Biology and Therapy Research Laboratory discovered that pinworm-infected mice, which were treated with mebendazole, did not develop appreciable brain tumors, even though the researchers had implanted brain cancer cells weeks before. In subsequent experiments, his team demonstrated that administration of mebendazole prevented tumor proliferation and improved survival times in mice by an average of 63 percent. Mebendazole was part of a phase I clinical trial for patients with newly diagnosed, high-grade glioma and glioblastoma.
This Phase 1 trial was primarily devoted to determining the safety and preliminary efficacy of mebendazole in treating glioblastoma. It has recently been completed – with encouraging results. Based on the promise of these early results, ABC2 is now working with Dr. Riggins to move mebendazole into Phase 2 clinical trials designed to more rigorously test its efficacy as a potential new glioblastoma therapy. If this project is successful, mebendazole could provide a safe, inexpensive, effective glioblastoma treatment with the potential for rapid translation into the clinic. Because mebendozole has been used around the world for years to treat pinworm infections in children, it offers a particularly enticing opportunity as a treatment for pediatric brain tumors.Recently, Bai and colleagues demonstrated compelling preclinical evidence for using the microtubule inhibitory drug mebendazole (MBZ) to treat several molecular subtypes of medulloblastoma, including group 3. As a long-standing antihelminthic drug, MBZ has the advantage of a low-toxicity profile in children compared with other microtubule inhibitors such as vincristine and paclitaxel. As a lipophilic agent with a low molecular weight, MBZ has the additional advantage of blood-brain barrier permeability. Previous studies suggest that MBZ acts as an inhibitor of vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2), the primary receptor mediating the effects of VEGF. This study reveals the antiangiogenic effect of MBZ in medulloblastoma preclinical mouse models and its encouraging impact on overall survival.  In another study Dr. Symons and colleagues examined mebendazole, a medication that is used to treat parasitic pinworms and that in previous studies had been found to be effective in the treatment of glioma tumors. By studying how mebendazole kills isolated tumor cells in the laboratory, they showed that it works in exactly the same way as vincristine. They also found however, that while mebendazole effectively slowed down the growth of glioma tumors, vincristine did not work at all."We were rather surprised to see that vincristine, which is currently used to treat a range of different brain tumors, was totally ineffective in our in vivo glioma model," said Dr. Symons. "In contrast, in the same model, mebendazole performed quite well, most likely because mebendazole crosses the blood-brain barrier and reaches the tumor much better than vincristine. The reason that vincristine may be erroneously believed to be effective for the treatment of brain tumors is that it always has been used in combination with other treatments."Based on the new results -- and due to the fact that vincristine often has severe side effects in comparison to relatively mild reactions to mebendzole -- Dr. Symons and his team are now strongly motivated to initiate clinical trials to test whether vincristine can be exchanged by mebendazole in the treatment of brain tumors."Sometimes innovation can be looking at an existing treatment in a new light," said Kevin J. Tracey, MD, president and CEO of the Feinstein Institute. "This new approach needs to be tested in clinical trials, but with Dr. Symons' new findings we may be closer to a new treatment option that could prolong the lives of the patients suffering from low-grade glioma and other brain tumors." Another study by deWitt et al also came to the same conclusion finding Vincristine ineffective and mebendazole quite effective in various brain tumors.

5. Zardaverine + Quazinone: Nadir M.
showed  that  subgroups  of  tumors,  within  many  different  cancer types,  overexpress  PDE3A  (mRNA  and  protein)  and  that  PDE3A  expression can predict sensitivity  to phosphodiesterase inhibitors.
Searching  the  Human  Protein  Atlas  database  revealed  that  differential PDE3A  expression,  as  observed  in  ovarian  cancer  specimens,  is  also  prevalent  in  many  other  cancer  types,  including  colorectal,  melanoma,  endometrial,  testis  and  urothelial  cancers. Their observations  suggest  that  PDE3A has  the  potential  to  be  both  a  biomarker  of  PDE  sensitivity  and  drug  target for cancer treatment.

6. Nelfinavir: earlier generation HIV drug with 15 years of safe use data has now been found to have multiple actions against cancer. It's action as monodrug is not as effective as an adjunct. Quoting a review article by Tomas K. "If apoptosis is described as a cascade, then apoptosis stimulator drugs like NFV should be viewed as enhancers of this cascade. An initiator of the cascade is still necessary, for example chemoradiotherapy. After this initial step, apoptosis stimulator drugs increase the amount of cells entering this pathway. This might be one of possible reasons why nelfinavir alone has shown poor results in a clinical trial used as monotherapy.

This does not mean that NFV cannot act as an initiator, but the evidences show that it is prone to be an enhancer of apoptosis rather than an initiator."

This review article also lists all the preclinical trials (Table 2) and clinical trials of Nelfinavir (Table 3):

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4457118/

7. Saquinavir-NO: This is another protease inhibitor like Nelfinavir but may be considered to have equal if not better action against cancers. Listed below are studies which demonstrate it's use and benefit:

ww.ncbi.nlm.nih.gov/m/pubmed/21170266/

https://www.ncbi.nlm.nih.gov/m/pubmed/21270522/

https://www.spandidos-publications.com/10.3892/or.2012.1840

8. CH05-10: An analog of Indinavir another PI has also shown broad spectrum anti cancer activity:

https://www.ncbi.nlm.nih.gov/m/pubmed/20946116/

9. Thalidomide + Lenalidomide:  Thalidomide was originally developed to treat morning sickness in pregnant women. Its use was stopped because it was found to cause birth defects. Thalidomide is now used as a treatment for cancer, but it must not be taken in pregnancy. A pregnancy prevention programme must be followed during treatment. Thalidomide works in several different ways. It helps the immune system attack and destroy cancer cells. Kill or stop the growth of cancer cells. It affects the chemical messages that cancer cells need to survive. It blocks the development of new blood vessels which cancer cells need to grow and spread. Thalidomide is usually taken with other chemotherapy drugs and steroids as treatment for myeloma. The researchers at Dana Farber Cancer Institute demonstrated that lenalidomide — a more powerful derivative of thalidomide — killed multiple myeloma cells by disabling overactive switches called transcription factors that drive the cells' excessive growth. Transcription factors are proteins that bind to genes and increase their activity, and cancers are often driven by overactivity of these molecular switches. For example, a transcription factor called c-Myc appears to be overactive in many different types of cancer.

10. DMOG dimethyloxalylglycine:
A team of researchers from the U.K. and the U.S. has found that a drug used to study hypoxia can also be used to inhibit glutamine metabolism—a possible means for targeting cancer cells by cutting off their supply line. In their paper published in the journal Nature Chemical Biology. The researchers began their study by noting that a lot of tumors require glutamine to survive—they utilize it in a process called glutaminolysis.  A drug called dimethyloxalylglycine (DMOG) might be useful in inhibiting glutamine used by cancer cells. Many types of cancer cells exhibit glutamine addiction. The growing cancer must synthesize nitrogenous compounds in the form of nucleotides and NEAAs. Glutamine is the obligate nitrogen donor. Glutamine’s contribution to amino acid biosynthesis establishes it as a key ingredient for the protein translation needs of cancer cells. A further role for glutamine in cancer cell protein translation stems from observations that a master regulator of protein translation, the mammalian target of rapamycin complex 1 (mTORC1), is responsive to glutamine levels. Glutamine consumption rate of many of the cancer cell lines exceeded the consumption of any other amino acid by ten-fold. Many cancer cell lines could not proliferate in the absence of exogenous glutamine and many could not maintain their viability in the absence of glutamine. Replenishment of the mitochondrial carbon pool by glutamine provides the mitochondria with precursors for the maintenance of mitochondrial membrane potential and for the synthesis of nucleotides, proteins, and lipids. 

wide variety of human cancer cell lines have shown sensitivity to glutamine starvation, including those derived from pancreatic cancer, glioblastoma multiforme, acute myelogenous leukemia, colon cancer and small cell lung cancer.

High Throughput Screening Robot

11. Blocking Cancer Exosomes:
Tulane University scientists Abdel Mageed, Amrita Dutta, et al used robotic high throughput screening technique to identify approved compounds that could block cancer exosomes. Apparently cancer exosomes are implicated in the spread of cancer. So any drug that blocks cancer exosomes from bring released by cancer cells can be very valuable to cancer patients at all stages. It can also be taken as an adjunct. They screened 4,580 compounds. The lead compounds tipifarnib, neticonazole, climbazole, ketoconazole, and triademenol were validated as potent inhibitors. 

12. Slowing down Cancer Metastasis:
I am a fan of young Johns Hopkins scientists Hasini Jayatilika. She and her colleagues discovered that it is not tumour size that triggers metastsis (parts breaking out to form tumours at other site) but it is tumour density that triggers metastasis. I reported about her significant discovery in one of my earlier posts on this blog. 
After 7 years of research she demonstrated how after reaching a certain density tumour cells begin to release two proteins Interleukin 6 and Interleukin 8. They tell the new cancer cells that it is getting too crowded separate out and build your own nest at some other site. She observed this after studying the communications between cancer cells. She also discovered two approved drugs known to work on Interleukin receptors that significantly slowed down metastasis:
Tocilizumab a rheumatoid arthritis treatment and Reparixin a potential cancer drug. When combined they seemed to significantly slow down the metastasis.

13. Cimetidine:

It was originally approved/used as a histamine blocker to reduce stomach acid secretion. But since many years multiple actions against cancer has been discovered. One of them is consistent with its previous use: some cancers release a lot of histamine to suppress immune response. Another one is even more startling: cancer cells have ligands Lewis X and Lewis A4 that bind to E-Selectin on endothelial walls for its metastasis. 'Since cimetidine inhibits the expression of E-selectin in blood vessels, cancer cells that are in the bloodstream can't bind to the blood vessels and establish a metastatic tumor. Instead they are eventually eliminated. This would obviously lead to a much better outcome for the patient. Indeed, patients with aggressive colon cancer (Dukes grade C) had a remarkable 84.6% ten year survival rate when treated with cimetidine for one year after surgery compared to a 23.1% ten year survival rate for patients that were not treated with cimetidine as an adjuvant therapy.'

Dr. Michele Morrow has written an article on Cimetidine's various actions against certain cancers like colorectal, gasttic, breast and pancreatic here:

https://www.lifeextension.com/magazine/2002/7/cover_cimetidine/Page-01

14. Clarithromycin:

Dr. Ferreri of San Raffaele Scientific Institute found the following in a human trial of 23 patients with high dose Clarithromycin:

Clarithromycin displays immunomodulatory and antineoplastic properties. As single agent, this macrolide is associated with tumor responses in anecdotal cases of relapsed/refractory extranodal marginal zone lymphoma (rrEMZL). Twenty-three patients were registered (median age 70 years, range 47–88 years; M:F ratio: 0.27) Tolerability was excellent, even among HBV/HCV-positive patients; only two patients had grade >2 toxicity (nausea). Six patients achieved a complete remission and six a partial response (ORR = 52%; 95% confidence interval 32% to 72%). Age, previous treatment and stage did not influence activity. At a median follow-up of 24 (16–33) months, only two patients with responsive disease experienced relapse, with a 2-year progression-free survival of 56 ± 10%; all patients are alive. Gauthier Bouche et al in a review article identified multiple myeloma, lymphoma, chronic myeloid leukaemia (CML), and lung cancer having the highest level of evidence of benefit of Clarithromycin.

15. ReDO Project:

In an article published in ecancer medical sciences by Gauthier Bouche and colleagues they write about the ReDO project in which they use high throughput screening and data from clinical trials to identify approved drugs for repurposing in oncology. 

Please do visit their site to learn more. This is a valuable resource for cancer patients and they should take full advantage of it and bring it to the attention of their oncologist if they see any of the identified drugs showing benefit on their cancer. They have so far identified 250 such drugs and compounds out of which they are first focusing on 6 Mebendazole, Nitroglycerin, Cimetidine, Clarithromycin, Diclofenac and Itraconazole. Start with this article:

https://ecancer.org/journal/8/full/442-the-repurposing-drugs-in-oncology-redo-project.php
Then go toReDO website:
http://www.redo-project.org

16. Metformin:
Is a leading oral tablet prescribed to millions of type 2 diabetics over many decades. All cancer patients should consider starting Metformin immediately after diagnosis. Due to its healthspan extension ability all adults above 40 years should consider it as a preventive. It is under phase II and phase III trials for its anti cancer actions. One of its primary actions is inhibition of mTOR1.
The mTOR pathway plays a pivotal role in metabolism, growth and proliferation of cancer cell. Increased levels of circulating insulin/IGF1 and upregulation of insulin/IGF receptor signaling pathways were demonstrated to be involved in the formation of many types of cancer. Metformin was found to reduce insulin level, inhibit insulin/IGF signaling pathways, and modify cellular metabolism in normal and cancer cells. Jacek Kasznicki of Medical University of Lodz and colleagues have written a useful review article in ATM Journal.

      Cancer cell under electron microscope
With Syrosingopine:
In another exciting finding published in Cell in 2018 titled 'Lethal combination: Drug cocktail turns of juice to cancer cells' by University of Basel, they discovered that metformin and syrosingopine originally approved as a hypertensive drug made cancer cells die. The combination of the two drugs blocks a critical step in energy production thus leading to an energy shortage, which finally drives cancer cells to "suicide". Cancer cells have high energy demands due to their increased metabolic needs and rapid growth. A limiting factor in meeting this demand is the molecule NAD+, which is key for the conversion of nutrients into energy. "In order to keep the energy-generating machinery running, NAD+ must be continuously generated from NADH," explains Don Benjamin, first author of the study. "Interestingly, both metformin and syrosingopine prevent the regeneration of NAD+, but in two different ways." Many tumor cells shift their metabolism toward glycolysis, which means that they generate energy mainly via the breakdown of glucose to lactate. Since the accumulation of lactate leads to a blockade of the glycolytic pathway, cancer cells eliminate lactate by exporting it from the cell via specific transporters. "We have now discovered that syrosingopine efficiently blocks the two most important lactate transporters and thus, inhibits lactate export," says Benjamin. "High intracellular lactate concentrations, in turn, prevent NADH from being recycled into NAD+."

Because the anti-diabetes drug metformin blocks the second of the two cellular pathways for NAD+ regeneration, combined metformin-syrosingopine treatment results in complete loss of the cell's NAD+ recycling capacity. The depletion of NAD+ in turn leads to cell death, as the cancer cells are no longer able to produce sufficient energy. Thus, pharmacological inhibition of lactate transporters by syrosingopine or other similarly acting drugs can increase the anti-cancer efficacy of metformin and may prove a promising approach to fighting cancer. The former Basel-based company Ciba originally developed syrosingopine for the treatment of hypertension in 1958. The identification of syrosingopine as a dual inhibitor of the two main lactate transporters is an important discovery, as currently there is no pharmacological inhibitor available for one of these two transporters (MCT4). The potential application of syrosingopine in cancer therapy could trigger a second career for this old drug.

17. Trifluoperazine:

Is approved as an anti-psychotic agent.
Pulloski-Gross et al demonstrated in 2014 that trifluoperazine is responsible for reducing the angiogenic and invasive potential of aggressive cancer cells through dopamine receptor D2 to modulate the b-catenin pathway and propose that trifluoperazine may be used as an antimetastasis chemotherapeutic.
In a screening experiment conducted by Yeh CT et al to target Cancer Stem Cells Trifluoperazine stood out for its action especially along with conventional therapy to stop proliferation of CSCs. In another post on this blog I have mentioned about the danger of drug resistance and remission due to CSCs. The combination of trifluoperazine with either gefitinib or cisplatin overcame drug resistance in lung CSCs. Trifluoperazine inhibited the tumor growth and enhanced the inhibitory activity of gefitinib in lung cancer metastatic and orthotopic CSC animal models. In another in vitro and in vivo study by Kang et el Trifluoperazine potently suppresses proliferation, motility, and invasion of glioblastoma cells in vitro, and tumor growth in in vivoxenograft mouse model. In another study in 2017 by iang et al on two HCC lines they found that
apoptosis was increased and the ability of migration or invasion was found to be impaired by Trifluoperazine. FOXO1 which acts as tumor suppressor on HCC lines expression was increased. Trifluoperazine in vivo could effectively restrict the angiogenesis and tumor growth with reduced expression of VEGF, Bcl-2, and PCNA, and increased the nuclear localization of FOXO1, which indicated its antitumor role in HCC. Terifluoperazine has been recently reported to show a strong anticancer effect on lung cancer, hepatocellular carcinoma, and T-cell lymphoma. Feng et al in 2018 found Trifluoperazine effective in prolonging survival and inducing apoptosis in Triple Negative Breast Cancer that has metastasized to the brain.Chlorpromazine:
Is another anti psychotic drug that has shown action in glioblastoma and colorectal cancer cells.

18. Disulfiram + Copper:
Like Metformin and Syrosingopine every cancer patient should consider taking this combo. Disulfiram also branded as Antabuse is prescribed to deter alchololics from drinking. There has been an amazing case of stage IV breast cancer patient whose cancer at age 38 had spread to her bones. Her doctors stopped all cancer medicine and prescribed her with Disulfiram. She died 10 years later out of a inebriated fall from a window and not cancer. During autopsy they were shocked to find that all her cancer in her bones had melted away leaving very few cancer cells in her morrow. Dr. Jiri Bartek of Danish Cancer Society Research Center Copenhagen and his colleagues in some brilliant work deciphered how Disulfiram killed all types of cancer cells. They showed that disulfiram and its main metabolite, ditiocarb, forms a complex with copper that blocks the machinery that cells use to dispose of misfolded and unneeded proteins. Partly because of the resulting protein buildup, the cancer cells become stressed and die. Bartek’s team also solved another puzzle—why normal cells aren’t harmed by disulfiram, even when patients take it for years. For unclear reasons, the copper metabolite is 10 times more abundant in tumor tissue compared with other tissues, the group found. Bartek and collaborators are now launching trials to test a disulfiram-copper combo as a treatment for metastatic breast and colon cancers and for glioblastoma, a type of brain cancer. Finding a new use for an approved drug is appealing because the compound has already passed safety testing. However, “big pharma probably won’t be interested” in developing disulfiram for cancer because there’s no patent protection on the drug, Bartek says. Still, if the pending clinical trials provide convincing evidence, oncologists could go ahead and prescribe it anyway as an inexpensive treatment. In a study published in Acta Biomaterials Huacheng et al administered Loaded Disulfiram nanoparticle plus copper ions (LDNP/Cu) wherein LDNP is DSF nanoparticle with lactobionic acid, a selective ligand for D-galactose receptor that is effective in targeting cancer cells to one group of mice with metastatic ovarian cancer from human cancer line. Once a week for 3 weeks. After monitoring tumor growth weekly, He and colleagues found that LDNP/Cu was the most effective in impeding tumor growth, and that rapid tumor growth occurred in controls. In addition, the researchers observed numerous tumors in the abdominal cavities of controls but fewer in mice treated with nanoparticles. Researchers evaluated the systemic toxicity of the nanoparticles, in part, via histologic exam of the liver, and found no noticeable differences between the treatment groups. “Altogether, the in vivo data indicate that the LDNP/Cu nanoparticle could be an effective and safe tool for the treatment of advanced ovarian cancer,” the authors concluded. In another study by Lui P et al Disulfiram abolished Cancer Stem Cell characters and completely reversed Paclitaxel and Cisplatin resistance in Triple Negative Breast Cancer cells. Young Min Park et al had success in remarkably slowing down head and neck squamous carcinoma cancer cell lines (FaDu and Hep2) in xenograft mice with Disulfirma and Copper.

19. Fenofibrate:
Since its clinical introduction as a third-generation fibrate in 1975, fenofibrate has been widely used in the treatment of hypercholesterolemia and hyperlipidemia. The lipid-lowering effect of fenofibrate is believed to be mediated through its stimulation of peroxisome proliferator-activated receptor α (PPARα). In addition to its lipid-lowering function, fenofibrate exerts also pleiotropic effects. For instance, fenofibrate was found to not only slow the progression of diabetic retinopathy and other microvascular complications in patients with type 2 diabetes, but also protect against retinopathy, nephropathy, and cardiac pathological changes in type 1 diabetes. Fenofibrate was established to afford myocardial protecttion through its direct effects on the cardiovascular system. Most recently, PPARα-specific agonists were reported to have anticancer effects in a large number of human cancer types, such as acute myeloid leukemia, chronic lymphocytic leukemia, and solid tumors, including those of the liver, ovary, breast, skin, and lungs. Furthermore, fenofibrate inhibited the proliferation of cell lines derived from breast and oral tumors, melanoma, lung carcinoma, glioblastoma, and fibrosarcoma in mouse models. The above is part of an excellent review article by Xin Lian et al published in 2018 in the Journal of Cancer. Here is link you can share with your oncologist:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5950581/

20. Flunarizine:
A drug approved for migraine has shown anti cancer actions in various studies. Dr. Schmeel et al of Center for Integrated Oncology, Bonn in a study had the following results: Flunarizine induced significant apoptotic activity in all tested myeloma and lymphoma cell lines in a dose-dependent manner. Conclusion: Our results reveal a significant selective induction of apoptosis by flunarizine and suggest an in vivo effect against lymphoma and myeloma. It was also found to positively modulate doxorubicin resistance in human colon adenocarcinoma multi drug resisting cells. Researchers at Baylor College of Medicine knew that a protein called Ras were drivers of wide number of cancers. They screened FDA approved drugs that could degrade Ras. Of all the ones they tested Flunarizine showed the best results. In their study published in Scientific Reports in 2018 Chang et al the researchers also tested the effect of flunarizine in a mouse model of triple negative breast cancer and found that it slowed down tumor growth. They also determined that flunarizine promotes N-Ras degradation by enhancing a natural cellular pathway called autophagy.

21. Ribavirin:
A broad spectrum anti-viral drug has shown action against multiple cancers. In a review paper by Dr. Katherine Borden with co-authors in 2010 concludes: Ribavirin targets at least two distinct biochemical entities: eIF4E and IMPDH. Ribavirin clearly targets the oncogenic activity of eIF4E in cell lines, in animal models, and in AML patients. Ribavirin monotherapy led to objective clinical benefit in many of these patients. The effects of ribavirin on the immune system, and whether these are mediated through eIF4E and/or IMPDH, may also play a role in its anti-cancer activities in AML patients. Given that eIF4E is up-regulated in over 30% of cancers, the hope is that ribavirin will become an important component in a wide variety of treatment regimens. 
Here is a link to the paper:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2950216/#__ffn_sectitle
In a more recent study of 2017 in xenograft mice model Shen et al showed that that ribavirin inhibited proliferation and induced apoptosis in the thyroid cancer cell lines 8505C and FTC-133. Ribavirin inhibited thyroid cancer growth in a xenograft mouse model. Ribavirin also sensitized thyroid cancer's response to paclitaxel. They concluded that their data clearly demonstrate that ribavirin acts on thyroid cancer cells by inhibiting eIF4E/β-catenin signaling. Our findings suggest that ribavirin has the potential to be repurposed for thyroid cancer treatment and also highlight the therapeutic value of inhibiting eIF4E-β-catenin in thyroid cancer. In another study Ribavirin showed benefit only in some cancers. Researchers investigated the growth inhibitory effects of ribavirin, the cell lines were exposed to different concentrations of ribavirin (10–50 μM) and cell viability was analyzed at 72 h. The results showed that the cell lines were inhibited in a dose-dependent manner although the extent of inhibition was cell line-dependent. The MCF-7 and MDA-436 breast cancer, DU145 prostate carcinoma and D54 glioma cell lines showed increased inhibition whereas the SW480 colon cancer, prostate PC3 and breast MDA-231 cells were less inhibited. HeLa cells showed minimal inhibition even at the highest dose of ribavirin.

22. SPHINX31:
In a new study (2018), Sanger Institute researchers and their collaborators set out to work out how inhibition of SRPK1 gene can kill AML cells and whether it has therapeutic potential in this disease. They first showed that genetic disruption of SRPK1 stopped the growth of MLL-rearranged AML cells and then went on to study the compound SPHINX31, an inhibitor of SRPK1, which was being used to develop an eye drop treatment for retinal neovascular disease – the growth of new blood vessels on the retinal surface that bleed spontaneously and cause vision loss. The team found that the compound strongly inhibited the growth of several MLL-rearranged AML cell lines, but did not inhibit the growth of normal blood stem cells. They then transplanted patient-derived human AML cells into immunocompromised mice and treated them with the compound. Strikingly, the growth of AML cells was strongly inhibited and the mice did not show any noticeable side effects.                                                                                                                                             23. Tubeimoside I (TBM):              Tubeimoside I (TBM) is extracted from the tuber of Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae), a traditional Chinese herb previously used in anti-viral or anti-inflammatory treatment.Growing studies have reported its direct cytotoxity in multiple human cancer cells, characterized by mitochondrial damage, endoplasmic reticulum stress, apoptosis and cell cycle arrest. In addition, TBM could sensitize human ovarian cancer cells to cisplatin (CDDP). TBM has been considered as a promising anticancer agent. Cerevical cancer is one of the most aggressive human cancers with poor prognosis due to constant chemoresistance and repeated relapse. Tubeimoside I (TBM) has been identified as a potent antitumor agent that inhibits cancer cell proliferation by triggering apoptosis and inducing cell cycle arrest. In a study by Xuping Fen et al in Cell Death and Disease a Nature publication found that TBM could induce proliferation inhibition and cell death in cervical cancer cells both in vitro and in vivo. Further results demonstrated that treatment with TBM could induce autophagosome accumulation, which was important to TBM against cervical cancer cells. Mechanism studies showed that TBM increased autophagosome by two pathways: First, TBM could initiate autophagy by activating AMPK that would lead to stabilization of the Beclin1-Vps34 complex via dissociating Bcl-2 from Beclin1; Second, TBM could impair lysosomal cathepsin activity and block autophagic flux, leading to accumulation of impaired autophagolysosomes. In line with this, inhibition of autophagy initiation attenuated TBM-induced cell death, whereas autophagic flux inhibition could exacerbated the cytotoxic activity of TBM in cervical cancer cells. Strikingly, as a novel lethal impaired autophagolysosome inducer, TBM might enhance the therapeutic effects of chemotherapeutic drugs towards cervical cancer, such as cisplatin and paclitaxel.                                                                                                                  24. Celecoxib + Digoxin:                                     I usually do not like to list anything unless some in vivo results are encouraging. But for this one I have made an exception because the logic sounds viable even in vivo and also because both the drugs are FDA approved their safety has been thoroughly tested. Of course the doctor would make the final decision. Vadim Backman et al at Northwestern discovered the changes of chromatin in cancer cells thanks an imaging technology developed by them called PWS microscopy. Complex
diseases such as cancer, Blackman says do not depend on the behavior of individual genes, but on the complex interplay among tens of thousands of genes. They used PWS to monitor chromatin in cultured cancer cells. They found that chromatin has a specific "packing density" associated with gene expression that helps cancer cells to evade treatments. The analysis revealed that a more heterogeneous and disordered chromatin packing density was related to greater cancer cell survival in response to chemotherapy. A more conservative and ordered packing density, however, was linked to greater cancer cell death in response to chemotherapy. "Just by looking at the cell's chromatin structure, we could predict whether or not it would survive," says Backman. "Cells with normal chromatin structures die because they can't respond; they can't explore their genome in search of resistance. They can't develop resistance." Based on their discovery, the researchers hypothesized that altering the structure of chromatin to make it more orderly could be one way of boosting cancer cells' vulnerability to treatment.On further investigation, the team found that they could modify chromatin's structure by altering electrolytes in the nucleus of cancer cells. The team tested this strategy using two drugs that are already approved by the Food and Drug Administration (FDA): Celecoxib and Digoxin. Celecoxib is currently used for pain relief, while Digoxin is used to treat atrial fibrillation and heart failure. Both drugs, however, are also able to change the packing density of chromatin. The researchers combined these drugs — which they refer to as chromatin protection therapeutics (CPTs) — with chemotherapyand tested them on cancer cells in the laboratory. According to Backman, they witnessed "something remarkable." "Within 2 or 3 days, nearly every single cancer cell died because they could not respond. The CPT compounds don't kill the cells; they restructure the chromatin. If you block the cells' ability to evolve and to adapt, that's their Achilles' heel." Although they say that one has not yet seen whether it works in live environment in vivo but all the cancer cell lines they tried it worked.                                                                                                              25. Diphenyleneiodonium:                         DPL can inhibit the production of vitamins that feed cancer cells, causing the cells to starve.“Our observation is that DPI is selectively attacking the cancer stem cells, by effectively creating a vitamin deficiency,” said researcher Michael Lisanti, MD, PhD. “In other words, by turning off energy production in cancer stem cells, we are creating a process of hibernation.” DPI stops the reproduction of cancer cells by cutting off their energy source, according to the study. The drug does this without creating the toxic adverse effects that are common with traditional chemotherapies, according to the study. The authors found that DPI had this effect on cancer stem cells, preventing the creation of more cancer cells. When added to a mixed population of cells, DPI sent stem cells into hibernation, according to the study; however, the drug did not fight against “bulk” cancer cells, which do not typically initiate tumor growth within patients, according to the authors. DPI was observed to inhibit more than 90 proteins enzymes from being converted into cellular energy in the mitochondria, according to the study. The lack of energy production deprives cancer cells of vitamin B-12 and riboflavin, which shuts the stem cell down and prevents growth. “It’s extraordinary; the cells just sit there as if in a state of suspended animation,” Dr Lisanti said. The treatment also works by weakening the cell and making it vulnerable to other drugs used to fight cancer, according to Dr Lisanti. “The beauty of this is that DPI makes the cancer stem cells metabolically-inflexible, so they will be highly susceptible to a many other drugs,” he said.

Cancer Stem Cells