After a great deal of early promise, resveratrol has been on the ropes for a while, most prominently as a result of studies questioning whether it can directly activate sirtuins — this against a backdrop of growing skepticism that sirtuin activation can extend mammalian lifespan in any case.

Now, another (possible) black eye: GlaxoSmithKline (the company that purchased Sirtris, a pharmaceutical company co-founded by sirtuin/resveratrol pioneer David Sinclair) has suspended a trial of a resveratrol formulation, SRT501 in multiple myeloma patients, because several of the study’s subjects developed kidney failure.

GSK emphasizes that the trial has not been cancelled, but they are observing a moratorium on recruiting new patients until they determine whether the resveratrol was responsible for the subjects’ kidney problems. Nephropathy is a frequent complication in myeloma; one hypothesis being entertained is that the very high doses of resveratrol used in the trial caused vomiting, which in turn resulted in dehydration and tipped the balance in kidneys already close to failure due to the underlying cancer.

More elsewhere:

Yesterday we learned that the most well-characterized mammalian sirtuin, SIRT1, is involved in the control of behavior in response to food availability. SIRT1 is just one of seven sirtuins in mammalian genomes, each of which has a characteristic expression pattern, subcellular localization, and physiological importance.

Today we’re going to talk about another member of the sirtuin family, SIRT3, which has been known for a while now to localize to the mitochondria. Now, Kim et al. have shown that the SIRT3 protein acts as a tumor suppressor:

SIRT3 Is a Mitochondria-Localized Tumor Suppressor Required for Maintenance of Mitochondrial Integrity and Metabolism during Stress

The sirtuin gene family (SIRT) is hypothesized to regulate the aging process and play a role in cellular repair. This work demonstrates that SIRT3−/− mouse embryonic fibroblasts (MEFs) exhibit abnormal mitochondrial physiology as well as increases in stress-induced superoxide levels and genomic instability. Expression of a single oncogene (Myc or Ras) in SIRT3−/− MEFs results in in vitro transformation and altered intracellular metabolism. Superoxide dismutase prevents transformation by a single oncogene in SIRT3−/− MEFs and reverses the tumor-permissive phenotype as well as stress-induced genomic instability. In addition, SIRT3−/− mice develop ER/PR-positive mammary tumors. Finally, human breast and other human cancer specimens exhibit reduced SIRT3 levels. These results identify SIRT3 as a genomically expressed, mitochondria-localized tumor suppressor.

So, the absence of the SIRT3 gene disrupts mitochondrial function and destabilizes the mitochondrial genome, but the causal relationship is unclear: one can imagine a derangement of mitochondrial morphology or physiology causing the genomic instability, but one can also imagine a primary defect in DNA metabolism causing the physiological defects — and one could also imagine both, operating in a vicious cycle.

The significance of the “single oncogene” observation requires a bit of explanation: In a normal cell, simply turning on a single tumor-promoting gene isn’t sufficient to transform the cell, i.e., create a tumor — if a cell detects a hyperphysiological level of a growth factor or internally generated mitogenic signal, it will undergo senescence, a permanent cell cycle arrest that (among other things) prevents mutated cells from turning into cancers.

In the case of the SIRT3 knockout, however, a single oncogene is enough to transform an otherwise normal cell, i.e., the loss of SIRT3 function appears to potentiate the transformation process. Furthermore, this implies that the mitochondria are involved in the decision to undergo senescence. This is not to say that SIRT3 is directly involved in the senescence fate decision: the SIRT3 knockout has broadly deranged mitochondria, and it’s not clear which mitochondrial function is involved in senescence.

While this report does not fully elucidate the mechanism of SIRT3 action, it is clear that oxidation must be central to the issue: reactive oxygen species (ROS) levels are high in the SIRT3 knockout, but increasing the dose of an antioxidant enzyme prevents the single-oncogene transformation. We don’t yet know whether the ROS themselves are causing oncogenic mutations, or the loss of the mitochondrial function in senescence is allowing cells that would have arrested to progress further down the path to cancer. (Could be both, obviously, but it’s likely that one of the two is more important.)

These findings are consistent with what we already know about the connection between mitochondrial ROS, cellular damage and lifespan (ideas that are central to one strategy for developing longevity-enhancing drug compounds by targeting antioxidants to mitochondria), but they raise a question: how much of the “longevity regulation” we owe to antioxidant genes can be attributed to tumor suppression?

ResearchBlogging.orgKim, H., Patel, K., Muldoon-Jacobs, K., Bisht, K., Aykin-Burns, N., Pennington, J., van der Meer, R., Nguyen, P., Savage, J., & Owens, K. (2010). SIRT3 Is a Mitochondria-Localized Tumor Suppressor Required for Maintenance of Mitochondrial Integrity and Metabolism during Stress Cancer Cell, 17 (1), 41-52 DOI: 10.1016/j.ccr.2009.11.023

There’s lots more about SIRT3 in a recent post at @ging.

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A review I should have cited yesterday addresses the promise and challenges of the two most prominent natural-product candidates for longevity-enhancing therapeutics. The author is prominent biogerontologist and all-around bright feller Matt Kaeberlein (see here for earlier posts on his group’s work).

Resveratrol and rapamycin: are they anti-aging drugs?

Studies of the basic biology of aging have advanced to the point where anti-aging interventions, identified from experiments in model organisms, are beginning to be tested in people. Resveratrol and rapamycin, two compounds that target conserved longevity pathways and may mimic some aspects of dietary restriction, represent the first such interventions. Both compounds have been reported to slow aging in yeast and invertebrate species, and rapamycin has also recently been found to increase life span in rodents. In addition, both compounds also show impressive effects in rodent models of age-associated diseases. Clinical trials are underway to assess whether resveratrol is useful as an anti-cancer treatment, and rapamycin is already approved for use in human patients. Compounds such as these, identified from longevity studies in model organisms, hold great promise as therapies to target multiple age-related diseases by modulating the molecular causes of aging.

Note that resveratrol has been taking a bit of a thrashing of late, with recently released studies calling into question its ability to directly activate sirtuins. Briefly, the critics posit that the early data may have been misinterpreted due to artifacts in a fluorescence-based system used to detect protein-drug interactions — but check comment #32 on that post for David Sinclair’s personal response on this issue.

ResearchBlogging.orgKaeberlein, M. (2010). Resveratrol and rapamycin: are they anti-aging drugs? BioEssays, 32 (2), 96-99 DOI: 10.1002/bies.200900171

I’m sitting in the Drexler Auditorium of the Buck Institute, where I’ve been working over the last six months. Today we’re being treated to an off-schedule “Special Institute Lecture” by Harvard’s Gregory Verdine. These are my notes about the talk; below, I’m paraphrasing Verdine’s words, not writing my own.

To fix it you have to find it: Repairing oxidative damage to DNA.

Mitochondrial metabolism generates oxygen radicals, which damage DNA and increase the risk of mutation. The primary oxidative adduct of guanine, 8-oxoG, differs by only two atoms from the original guanine, but this small difference is still enough to change the residue’s base-pairing characteristics (i.e., from G=C to 8-oxoG=A). 8-oxoG is repaired by the enzyme Ogg1, which displaces the damaged residue by covalently bonding to the DNA backbone.

Ogg1 has a tough job: oxoG-C base pairs are perfectly stable, and from the outside they look just like G-C pairs, so the detection of rare lesions within the genome poses a tremendous challenge. In crystallographic studies that exploited a catalytically dead Ogg1 enzyme (which can recognize but not cleave DNA), Verdine’s lab has shown that the binding of Ogg1 at a G-C base pair results in the extrusion of G or oxoG to the exterior of the double helix, setting the stage for repair — but how does Ogg1 find the lesion site in the first place? To answer this question, Verdine’s group visualized single molecules of Ogg1 diffusing in one dimension along a double helix. They observed that Ogg1 moves so quickly that it can’t be checking every G-C base pair along its path. Instead, Ogg1 (and other lesion-repair proteins) may exploit subtle rearrangements in the DNA backbones near lesion sites. The enzyme amplifies these local structural changes into substantial conformational changes, leading to base extrusion and starting the process of repair.

We want a new drug: Synthetic biologicals as novel pharmaceuticals

Small-molecule drugs have “good geography,” in the sense that they can cross cell membranes. However, they’re limited, in that they can only target proteins that engulf them (e.g., in hydrophobic pockets or active sites). Proteins, on the other hand, have much more diverse function, but terrible geography — they simply can’t get into cells, and they’re therefore useless for intracellular targets. Both classes of drug can attack (generously) only ~10% of prospective targets. Therefore, we need an entirely new class of drug: synthetic biologics like stapled peptides and RIPtides, which combine the bioavailability of small molecules with the functional diversity of proteins.

Stapled peptides are essentially the interaction domains of proteins, conformationally restrained in such a way that they still retain the active structure. (Think of a protein as a delivery system for an interaction domain, in which the non-interacting portions serve primarily to hold the ID in place.) An interaction domain alone would be too floppy to have a biological effect; conversely, the intact protein has the desired function but can’t cross the membrane. Solution: replace the main body of the protein with a hydrocarbon “staple” that keeps the interactive domain in the active conformation, without substantially increasing its size. Surprisingly, stapled peptides are taken up by cells via an energy-dependent active transport process, one upshot of which is that they don’t need to be uncharged and hydrophobic in order to cross the membrane. Drugs of this kind have already been used in animal studies to suppress leukemia by activating apoptotic factors in tumor cells.

Declaring open season on transcription factors

A brief concluding note: Transcription factors are among the most well-validated prospective targets, but they have historically been outside the scope of drug developers. TFs function primarily by protein-protein interactions that aren’t amenable to interference by small-molecule drugs. Recently, however, Verdine and others have been able to use synthetic biologicals to interfere with a specific oncogenic transcription factors.

My own comments

OK, so, not a lot of specific about either aging or cancer, but the idea of a novel class of pharmaceuticals that could be used to attack the “missing 80%” of validated prospective drug targets is still very exciting.

UCSC, the institution that brought you the industry-standard genome browser, has now launched the UCSC Cancer Genomics Browser:

The browser is a suite of web-based tools to integrate, visualize and analyze cancer genomics and clinical data. This browser displays a whole-genome and pathway-oriented view of genome-wide experimental measurements for individual and sets of samples alongside their associated clinical information.

This site hosts the public UCSC Cancer Genomics Browser. The public site contains a rapidly growing body of publicly available cancer genomic data, including 12 published studies, datasets from the TCGA consortium, and others.

We encourage you to explore these data with our tools. The browser enables investigators to order, filter, aggregate, classify and display data interactively based on any given feature set including clinical features, annotated biological pathways, and user-edited collections of genes. Standard statistical tools are integrated to provide quantitative analysis of whole genomic data or any of its subsets.

I suspect that the Cancer Genomics Browser will provide an indispensable tool for biogerontologists who are seeking to explore the mechanistic connections between aging and cancer. I’m currently trying to think up an interesting way to use the service (and publicly available data) in my own work: e.g., tumors all have to undergo cellular senescence; would it be possible to find some fingerprint of senescence bypass mechanisms by looking at expression data from large numbers of tumors?

Here’s the latest in our series of review roundups — links, without extensive further comment, to the reviews I found most intriguing over the past few weeks. For the previous foray into the secondary literature, see here.







Stem cells:


Here is the next in what will likely be a long series of semi-regular review roundups — links, without extensive further comment, to the reviews I found most intriguing over the past few weeks months (I went on hiatus during the winter holidays). For the previous foray into the secondary literature, see here.


Apoptosis & cancer:

Calorie restriction:




Stem cells:


I’m sitting in an auditorium listening to a seminar by Laura Niedernhoefer from U. Pittsburgh. She’s telling us that Ercc1-/- mice, which are deficient in nucleotide excision repair, show transcriptional changes that mirror those found in old wildtype or unusually long-lived mutant animals. Her data is strongly reminiscent of recent findings that progeroid DNA repair mutants exhibit transcriptional similarities to aging calorie-restricted and dwarf animals. As in the earlier studies, the Ercc1-/- animals showed a marked downregulation of the somatotroph (GH/IGF-I) axis — suggesting that persistent DNA damage causes the body to direct energy away toward repair and away from growth.

Curiously, the Ercc1-/- mice are resistant to both cancer initiation and tumor progression — they rarely form tumors of their own, and even whopping doses of aggressive fibrosarcoma cells (from ERCC1 wildtype animals) don’t result in significant tumor growth. Largely on the basis of the latter finding, Niedernhoefer attributes the decrease in cancer to the downregulation of somatotrophic signaling; the idea here is that tumors can’t get off the ground without the cocktail of growth factors present in an animal with a normal GH/IGF-I axis.

The final chapter of the talk dealt with the mechanism of the ERCC1/XPF nuclease (an obligate heterodimer of the two proteins). Knockout animals showed a specific defect in accurately resolved experimentally induced double-stranded breaks with 3′ overhangs — other sorts of DSBs were processed equally efficiently in wildtype and knockout cells. Genetic crosses with other DSB repair mutants demonstrate that ERCC1 nuclease acts at a different step than either DNA-PKCS or Ku, two other key factors in the cellular response to DSBs.

Niedernhoefer concluded her talk with the provocative idea that DNA cross-links in real cells (as opposed to cells in an experimental setting) are largely the consequence of molecular species formed by lipid peroxidation: reactive oxygen species (ROS) oxidize polyunsaturated membrane lipids, which then fragment into reasonably stable multifunctional unsaturated compounds that are much more stable than the parent ROS and therefore more likely to make it to the nucleus, where they can modify DNA to form cross-links. Consistent with this, diets high in polyunsaturated fats dramatically decrease the lifespan of ERCC1 mutants (which are sensitive to all kinds of DNA cross-links).

Many cells within a tumor are capable of initiating tumor formation on their own. This finding sticks a pin in the theory that tumorigenesis is driven and controlled by rare cancer stem cells — and suggests that therapeutic strategies targeted at cancer stem cells may be misguided. From Quintana et al:

Efficient tumour formation by single human melanoma cells

A fundamental question in cancer biology is whether cells with tumorigenic potential are common or rare within human cancers. Studies on diverse cancers, including melanoma, have indicated that only rare human cancer cells (0.1–0.0001%) form tumours when transplanted into non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. However, the extent to which NOD/SCID mice underestimate the frequency of tumorigenic human cancer cells has been uncertain. Here we show that modified xenotransplantation assay conditions, including the use of more highly immunocompromised NOD/SCID interleukin-2 receptor gamma chain null (Il2rg-/-) mice, can increase the detection of tumorigenic melanoma cells by several orders of magnitude. In limiting dilution assays, approximately 25% of unselected melanoma cells from 12 different patients, including cells from primary and metastatic melanomas obtained directly from patients, formed tumours under these more permissive conditions. In single-cell transplants, an average of 27% of unselected melanoma cells from four different patients formed tumours. Modifications to xenotransplantation assays can therefore dramatically increase the detectable frequency of tumorigenic cells, demonstrating that they are common in some human cancers.

(Hat tip to Longevity Meme)

The journal Cancer has a special supplemental issue on “Cancer and Aging: Challenges and Opportunities across the Cancer Control Continuum“.

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