Saturday, 14 December 2019



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:
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.

Saturday, 29 December 2018



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 vivoin 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.!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):

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:

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

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:

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:
Then go toReDO website:

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:

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:
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

Sunday, 24 June 2018


One more evidence

DKFZ The German Cancer Research Center is Germany's largest biomedical research institute with a thousand scientists. On 20th June 2018 they reported a discovery in the field of aging. I will quote some of what they report: 'Oxidative stress causes cells and entire organisms to age. If reactive oxygen species accumulate, this causes damage to the DNA as well as changes in the protein molecules and lipids in the cell. The cell ultimately loses its functionality and dies. Over time, the tissue suffers and the body ages. "The theory of oxidative stress or the accumulation of reactive oxygen species as the cause of aging has existed since the 1950s," says Peter Krammer of the German Cancer Research Center (DKFZ). "So far, however, the details of this process were unclear."
One way in which the body disposes of harmful reactive oxygen species is their conversion by the enzyme thioredoxin-1 (TRX-1). TRX-1 has been proven to play a role in protecting DNA from oxidative stress and slowing down aging processes. Its antagonist TXNIP inhibits thioredoxin-1 and thus ensures that the reactive oxygen molecules are retained.
In fact, reactive oxygen species (ROS) do more than just damage the body. For example, they are essential for the T-cells of the immune system to become active.' So ROS is also beneficial in the right proportion. The key is for excess ROS to be processed away before it can become harmful. But inversely, too much TRX-1 can impair critical immune related activity triggered by ROS. As I have stated in my earlier blog post called 'Mechanism of Aging' Nature modulates levels by using agonist and antagonists. It's like a hot water tap and a cold water tap and one can adjust their output to get water at just the right temperature. Where water represents repair pathway. Excess of either cold or hot can make it painful and harmful for us. Just the right temperature is called homeostatic state in our biology. 
When we are young the agonists Trx1 and antagonists TXNIP maintain just the right levels of ROS. But as per theory proposed in my earlier post ' Mechanism of Aging' Nature selectively increases the levels of antagonists as we age in a progressive manner. This begins to progressively increase the levels of ROS. To continue with my comparitive example it's like cold water is drowned out by increasing levels of hot water. At one point it begins to scald. That is what happens with uncontrolled ROS damaging for example DNA to a scale at which DNA's repair enzymes can't keep up. Nature by the way also modulates DNA repair systems with agonist and antagonist molecules. DNA repair is activated by PARP1 whose antagonist is DBC1 and agonist is NAD+. So can you see a pattern emerging here? TXNIP domination over TRX-1 as we age leads to rising ROS levels. One of the damage excessive ROS causes is to DNA. But as we age and just when DNA needs it most DBC1 levels also begins to dominate over NAD+ levels. So ROS causes more DNA damage just when less and less repair enzymes are available to repair it. Mounting unrepaired DNA damage then leads to rising mutations and genomic instability. 
I am excited by the DKFZ findings as it corroborates the theory proposed in mechanism of aging post on this blog of how aging program is implemented in our body. What you read above is happening in all major repair pathways and it snowballs progressively making death unavoidable. But what would happen if we selectively upregulate the dwindling agonists of repair? In the case of upregulation of NAD+ David Sinclair's lab at Harvard Medical School showed that DNA repair markers improved. Similarly the DKFZ researchers led by Krammer and Gülow wanted to know whether more TXNIP is formed in the body with increasing age, thereby undermining the protective mechanism against oxidative stress. To this end, they first compared T cells from the blood of a group of over 55-year-old volunteers with the T cells of younger blood donors, who were between 20 and 25 years old. In fact, it turned out that the cells of older subjects produce significantly more TXNIP. The DKFZ scientists have also observed similar findings in other human cell and tissue types. They also found TXNIP levels similarly much higher in older flies. Upregulation of TRX-1 would balance out the higher levels of TXNIP thereby forestalling the damaging chain reaction of rising ROS levels.
Another Evidence:
Autophagy is a very important repair and recycling pathway. With aging this too progressively goes down in efficiency leaving more and more senescent cells. It's importance can be gauged by the $307 million raised by Unity Bio based on pre-clinical data about a molecule that can clear senescent cells. One of their investors is now the world's wealthiest man Jeff Bezos founder of Amazon.
As cells die autophagy recycles the dead cells into new cells and clears debris. If this doesn't happen efficiently, aging humans are left with dead zombie cells (senescent cells) that secrete damaging cytokines harming surrounding healthy cells leading to mounting damage in tissues and organs.
Alvaro Fernandez and Salwa Sebti along with colleagues at University of Texas Southwestern Medical Center, USA published a study in Nature Journal on 30th May 2018. As per theory of mechanism of aging posted on this blog there is negative regulator of autophagy - antagonist called BCL2. It interacts with Beclin 1 a regulator of autophagy and apoptosis and blocks autophagy. As we age guess what happens. BCL2 increases it's interaction with Beclin 1. When the scientists disrupted this interaction between them it increased autophagy. Just bringing up one major repair and recycling pathway back to youthful levels not only increased healthspan but also lifespan. 
Now as promised I am sharing very good news for all who wished us luck for our pre-clinical trials mentioned in the post Mechanism of Aging: We have the results of a 2 month study where we selectively administered natural molecules and compounds that upregulate major known repair pathways. First of all safety tests cleared with flying colors histopathologist could not distinguish between young and treated old tissues. The main trial results were also spectacular: chronic inflammation of treated old rats reversed all the way back to levels of young control rats! Muscle strength - grip strength of old treated rats went up almost reaching young control rats! And cognitive skills - memory test scores of old treated rats were close to young control rats! The old treated rats lost weight despite having same amount of feed as old untreated rats! There was a wide difference between the levels of old treated rats and old untreated rats. If these are some of the key markers of aging did we reverse the age of old treated rats back to youth? It was equivalent of a 70 year old developing the metabolism, strength and memory of a 20 year old. Does it prove the theory of how Nature implements it's aging program? The anti aging benefit derived by rapamycin and metformin are limited and triggered only in a narrow range as it is a hormetic intervention. Anti aging benefits derived by resettng gene expression which has been methylated by aging changes to the epigenome theoretically may not be restricted by a narrow bandwidth. We have to find out over hundreds of studies whether this can be the fountain of youth we have been searching from thousands of years. Now on to human clinical trials. Please wish us luck.

Friday, 1 December 2017


Hormesis is evolutionarily conserved stress response. No other anti aging therapy has shown beneficial results in humans so far except Hormetic interventions like Calorie Restriction, prolonged fasting, intermittent low dose rapamycin and metformin. 
We come across all kinds of stress on a daily basis like UV radiation, very hot or cold exposure, prolonged hunger, high intensity exercise, etc. Our body has adaptive genes that activate boosters to cope with these minor stress events and minimize damage to the system. At a sweet spot of the amount of stress Hormesis leads to house cleaning with activation of autophagy and mitophagy. This recycles senescent cells and leads to mitogenesis. It also activates Nrf2 which releases enzymes that reduce the burden of reactive oxygen species ROS amongst 200 other beneficial gene expressions. It protects against age related chronic disorders including diabetes, middle aged obesity, cardiovascular disease, cancer and neurodegenerative diseases (Mattson, 2008). This post is not a detailed study of hormesis but a layman's update about benefits of hormesis and how one can take advantage today. So there are many other beneficial hormetic actions but the above examples illustrate their purpose. It launches a cascade of renewal including stem cell renewal, release of beneficial enzymes and hormones (for example an oxytocin bath) and clean up of wastes which allows the system to use it's full resources to recover from the stress. For a more comprehensive understanding of hormesis please read posts on the blog 'Anti Aging Firewalls' by Vincent Giuliano. You can also read 'Hormesis, Adaptive Epigenetic Reorganization, And Implications For Human Health and Longevity' by Alexander Vaiserman published in 2010 in Dose Response. Hormetic response is only available against stress up to a certain threshold with regards to intensity and time. It has a sweet spot or narrow  range where it leads to an over compensation. The beneficial cascade not only helps recover from the stress but also then goes ahead and delivers system wide improvements. This over compensation is what provides all the anti aging benefits of hormesis.  Hormesis can also be seen in plants. For example when we boil raw peanuts or tomatoes their nutritive value increases in multiples. Although limited the benefits can be potent. All system wide anti aging successes seen so far in humans use hormesis. For example Calorie Restriction at identified levels creates a stress which triggers a hormetic response leading to all its studied benefits. Valter Longo director of University of Southern California Longevity Institute has published an amazing study which concludes that prolonged fasting of 7 days triggers a stem cell based repair and regeneration of our hematopoietic system. Similarly rapamycin is a poison which targets mTOR. Its inhibition due to a low intermittent dose leads to partial reversal of the damage caused by aging. mTOR is inversely related to many pathways involved in renewal and repair like Nrf2, AMPK, etc. From a different path Metformin also generates hormetic benefits. Diabetic patients on Metformin it has been found in large studies tend to outlive patients on other medicines and also non diabetics. Even the most popular health intervention exercise provides it's most powerful benefits due to hormesis. 
So can hormesis stop aging and increase lifespan. No. As we age our repair and renewal machinery begins to lose its efficiency gradually in a progressive manner. Hormesis provides boost to the repair and renewal systems. Regular use of it would slow down the loss of efficiency thereby reducing unrepaired damage and build up of senescent cells, cancer causing mutations and protein aggregates. This has shown to improve healthspan and help prevent diseases like middle age obesity, metabolic syndrome, type 2 diabetes, cardiovascular disease, cancer, neurodegenerative disease, etc. This in turn may lead to reaching longer lifespan.  
I have started on rapamycin (Sirolimus) thanks to Dr. Alan Green (please web search for his name and rapamycin and it will lead to his website). On his website he has given his experience with rapamycin along with some very useful information about dosage, half life, strategy to avoid side effects, etc. He suggests 2 options conservative could be 3mg per 10 days. Aggressive would be 6mg per week. FDA states a elimination half life of 62 hours +/- 16 hours. So in a week it would be 2.7 half lives. If we take a shorter interval than it may lead to accumulation of the toxin which is not a good idea. Dr. Green as he says on his website ran a marathon in 4 hours at age 40 but by the time he reached 70 his physical activity was restricted to walking his dog. At age 72 he experienced angina and shortness of breath - his fasting blood sugar was high, creatinine was high and he could not fit in any of his pants. He discovered rapamycin and Koschei through Blagosklonny. He decided to take the aggressive approach. He shared that the results in 4 months were miraculous: he lost  20 pounds, his waist line went from 38 to 33 inches. He could walk 5 miles a day and ride bike over hills without angina. His blood sugar and creatinine went back to normal. He reported no side effects (no mouth sores). He felt from feeling old to feeling young. He has called rapamycin the world's greatest medicine. I have had a few exchange of messages with Dr. Green on an anti aging forum and can vouch that he is skeptical of all science unless it has shown replicable evidence. Someone younger may want to consider the conservative approach. Of course rapamycin and metformin both are prescription drugs and must be taken only after consulting your doctor. I personally started with 1mg and after 9 days taken 2mg and have now stabilized at 3mg. Also space weights training and other heavy exercise away from rapamycin days. It is important to do exercise as that too produces hormesis and allows us to make the most of taking rapamycin. Let me give you an example. In our middle age rogue macrophages block signal from our brain to our visceral fat cells to burn fat when we need energy for example during exercise. But when rapamycin benefits reconnect the signal it will not do any good unless we create an energy demand by doing intense exercise. With regards to Metformin one can create a hormetic effect every day whereas in the case of rapamycin there would be a tapering off between doses as part of the cycle. Not yet come across a study comparing the hormetic benefits between the two. Life Extension foundation has a good web page discussing metformin dosage for non diabetics at: 
They mention 500 mg twice a day for anti aging benefits for non diabetics but some scientists believe such benefits start only at 1,500 mg a day divided in 3 doses. To be noted that anyone having liver, kidney or congestive heart failure should not take this. One must also consider that no human trials have been done in either specifically for their anti aging benefits. That doesn't mean hormetic benefits are speculative but it leaves a gap with regards to ideal dosage for optimum benefits and safety. My personal opinion is not to take both rapamycin and metformin as too much of mTOR inhibition too is not good. When one does weights training one needs the growth with an activated mTOR. An article by Blagosklonny published in Aging 2010 cites two studies that show that inhibition of mTOR beyond a certain level causes depletion of male hormones which would adversely affect ones sex life. Therefore it may be better to have a cyclic intervention like rapamycin with a 10 day interval rather daily dose of metformin. One can investigate whether cyclic dosing of metformin provides similar benefits. I do take DHEA about 4 days away from rapamycin to make up for any deficiency in testosterone.  People as young as 35 are starting on one of the two to protect their future from age related diseases but one must avoid such mTOR inhibition in younger than 35. Many scientists wrongly believe that chronic inhibition of mTOR during aging leads to sustained anti aging benefits. Even when we are young mTOR swings between activation and inhibition based on its role as an important sensor. Insulin, cytokines, nutrients, and tesosterone stimulate cellular growth in part by activating the mTOR pathway. At puberty there seems to be optimum balance between mTOR activation for growth and mTOR inhibition for repair. As we age due to falling levels of repair the balance is lost which leads to over activation of mTOR. All we need is either intermittent activation of repair (a strategy my venture is pursuing) or intermittent inhibition of mTOR. Either action should lead to reduction of their imbalance. Which in turn should slow down the epigenetic drift - and in turn aging itself.
With regards to hormesis what is exciting though is that for the aging this system wide benefit is available today! One could begin to benefit immediately.
This is not a cure for aging or death but is still the most significant recourse available today for humans to improve old age. If we can slow down aging we may be alive when the cure for aging is discovered. In the future people will see old movies just to find out about the disease of old age and gasp at how horrible it made us look and feel. My venture to hunt for the cure just progressed further - We have a CTO who is a Professor of Natural Sciences from 23 years of a highly reputed USA University and we just signed a research collaboration agreement with a very highly reputed University for pre-clinical trials - 14 in all. We are awaiting Ethical Committee approval and hope to launch by first week of December. Wish us luck. We are developing nature or endogenous derived interventions that also inhibit mTOR like rapamycin and metformin but indirectly by upregulating or activating key repair pathways. It seems that Repair and activated mTOR are inversely related. Our interventions are expected to have amongst the highest upregulation of beneficial gene expressions available. On a separate note I was asked what will you look for if you were able to live longer. I said: Purity. The most prized human trait after unsolicited acts of kindness.