Telomeres and Telomerase: A Modern Fountain of Youth?

Interestingness: 3

By João Pedro de Magalhães and Olivier Toussaint, in Rejuvenation Research, July 2004, 7(2): 126-133. doi:10.1089/1549168041553044.

Not as much new material in this short paper as I expected, probably because I've read quite a bit on the topic since this was published. Interesting bits picked out:

• No correlation between maximum number of cell replications (cumulative population doublings) and age (post birth). (I have the vague recollection of some paper saying the opposite)
• No connection between mean telomere length and mammalian aging. (I think they mean across species, the species with the longer telomeres don't live any longer).
• Telomere length in vivo has very high variability

In summary, they think that telomerase might be beneficial for specific diseases but they doubt it'll turn out to be an anti-aging agent.

Telomere Length Dynamics in Normal and Malignant Hematopoiesis

Interestingness: 2

By Tim H Brümmendorf, Peter M Lansdorp and Nathalie Rufer, in the Journal of Anti-Aging Medicine, Volume 3, Issue 4, 2000 (pp 397-409, doi:10.1089/rej.1.2000.3.397.)

This seems to be a double-length episode of the previous paper, relating how telomere lengths change in blood cells across time and diseases. One thing it clarified was that they thought the shortening of telomeres in bone marrow donor recipients was due to the repopulation of the bone marrow stem cells from the few donor cells.  Also, they kept on a theme that the telomere lengths of most blood cells are merely reflecting the lengths in the haematopoietic stem cells (HSC aka bone marrow stem cells) which produced them.

Telomerase Activity and Telomere Length in the Haemopoietic System: Changes with Aging, Disease, and Therapy

Interestingness: 3

By JD Robertson and RF Wynn, in the Journal of Anti-Aging Medicine, Volume 3, Issue 4, 2000 (pp 389-395, doi:10.1089/rej.1.2000.3.389)

Review of what was known about telomere length and telomerase in blood cells. Haemopoietic stem cells (HSC) have active telomerase but their telomeres shorten gradually. Same for T-cells. Telomeres in neutrophils also shorten at about the same rate as in T-cells (20-50 base pairs per year), but from what I understood, they don't have active telomerase, so the telomerase is acting as a compensation method for the occasional clonal expansion of T-cells.

Checking for which X chromosome is inactivated, in old women most blood cells have the same one, as if they come from fewer and fewer stem cells.

In people with acute leukemia, telomeres are short and telomerase long, and they suggest this suggests that telomerase activates late in the process of disease. Also short telomeres on aplastic anemia and Fanconi's anemia.  Also shorter telomeres in bone marrow transplant recipients than in donors (about 15 years worth), but I'm not clear if they are saying this is because the transplanted tissue has had to undergo quick replication to refill the recipient's system, or that this was there before.

DNA Damage and Telomere Length in Human T Cells

Interestingness: 3

By Yvonne A Barnett, Christopher R Barnett, and Thomas Von Zglinicki, in the Journal of Anti-Aging Medicine, Volume 3, Issue 4, 2000 (pp 383-388, doi:10.1089/rej.1.2000.3.383. )

Seemed speculative on the link between T-cell, telomere length and oxidation damage in vivo, but presented a series of facts to suggest a link: increasing mutations in one particular gene in lymphocytes with age, senescent-looking T-cells in vivo in centenarians and people with Down's syndrome, and fibroblast with higher anti-oxidant capacity having a slower telomere shortening rate in vitro.

(Interesting: link between shorter telomeres and vascular dementia, but not with Alzheimer's, stroke or heart attack)

Cellular Senescence Mechanisms Independent of Telomere Shortening and Telomerase: Other Barriers to Cell Immortalization and Carcinogenesis

Interestingness: 4

By Izumi Horikawa, Toshio Yawata, and J Carl Barrett in the Journal of Anti-Aging Medicine, Volume 3, Issue 4, 2000 (pp 373-382, doi:10.1089/rej.1.2000.3.373.)

 Not all cells given telomerase escape senescence and cells with active telomerase can be made to senesce in lots of ways. The p16^INK4A/RB pathway can trigger senescence as well as the p14^ARF/MDM2/p53 pathway. p53 is probably related to telomeres but other parts probably aren't. By introducing single chromosomes into immortal cancer cell-lines and making them senesce, they infer the existence of other independent pathways of senescence. The mechanisms that trigger senescence are cell-type dependent.

Interesting factoid: mouse cells senesce after much fewer replications (10-20 vs 50-80) even though much longer telomeres.

Telomeres, Telomerase, and Premature Aging

Interestingness: 4

By Corrin V Wallis and Richard GA Faragher in the Journal of Anti-Aging Medicine, Volume 3, Issue 4, 2000.

Nice summary of telomeres, telomerase and also about the relations of telomeres, Werner's syndrome, Hutchinson-Gilford progeria syndrome and aging. Quite a few details of the proteins involved that I wasn't aware of and that I'll forget about in the next hour.

Is Telomere Shortening Related to Progeria?

Summary: Telomere shortening probably doesn't cause Hutchinson-Gilford progeria

Interestingness: 6

Paper by W Ted Brown in the Journal of Anti-Aging Medicine, Volume 2, Issue 2, Summer 1999.


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This is a short note speculating on whether progeria is caused by telomere shortening. The author says unlikely. Hutchinson-Gilford progeria is a fucking rare disease (1 in 8 million) of the type that pop up through a dominant spontaneous DNA mutation. Progerias are diseases that look like accelerated aging. This one starts being noticeable in toddlers between one and two years old, then they start looking old very quickly, going bald and losing subcutaneous fat, and have an expected lifespan of 13 years. 80% of them die of heart attacks and congestive heart failure, but they don't seem to get cancer, cataracts, osteoporosis or Alzheimer's like regular old people.

Fibroblast cultures extracted from progeria patients have an almost normal lifespan, but one paper reported shorter telomeres in them. Studies from Werner's syndrome, a different progeria that hits during early adulthood, give mixed results for shorter telomeres, but maybe some indication of faster telomere shortening.

Mice with telomerase knocked out don't show too many problems and in one study, could reproduce for at least six generations. By the sixth generation, their telomeres were much shorter and there were a lot of chromosome fusions. Other studies on these telomerase knockouts showed slightly lower lifespan, lower wound healing capacity, and more cancer. From this, he says it seems unlikely that telomere shortening would cause progeria. From what I remember, though, mice have way longer telomeres than humans to begin with, which would hide the effect a bit, but he didn't discuss that

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Importance of T-Cell Replicative Senescence for the Adoptive Immunotherapy of Cancer in Humans?

Summary: Review of replication of T-cells in vitro

Interestingness: 3

Paper by Graham Pawelec in the Journal of Anti-Aging Medicine, Volume 2, Issue 2, Summer 1999.

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This is another paper on T-cell senescence (previous one here: http://readingrejuvenationresearch.blogspot.com/2010/06/immunosenescence-analysis-and-genetic.html). It focuses on in-vitro studies, saying they are clinically important since that is how immune therapies will likely work best (eg training T-cells on tumor cells outside the body and then reinserting them) to work around the low immune responses of old people. I think their main area of investigation is trying to optimise the conditions under which T-cells replicate the longest.

It says the average number of population doublings (PD) of a T-cell in vitro before it becomes senescent, when externally stimulated, is 17, but 33 for cells that manage to get "established" (ie they get to a million cells). Seems like a very arbitrary cutoff but it better matches the numbers in the previous paper (25-40). The longest living ones reach 80 PDs on average and their record is around 170. They don't know why the large variability exists. The age of the person they were taking from doesn't seem to be one of the important variables. Longevity of CD34+ stem cells differentiated in vitro is no different to that of mature CD3+ cells.

They then switch to the link between telomeres and senescence. Fibroblast telomere length is directly proportional to replicative capacity. They say that this might apply to lymphocytes since the telomere lengths of human blood cells ex vivo are related to donor age, and the rate of telomere shortening with each doubling is about the same as for fibroblasts (120 bp per cell doubling). To me this would contradict what they said before that the replicative longevity was not related to the age of the donor, unless they mean blood cells other than T-cells.

In experiments by other people (Weng, Levine, June, et al) they found that CD4+ memory cells have shorter telomeres than naive cells, and that the difference is independent of the age of the donor. Telomere length decreases during autocrine replication of both of these and naive cells have higher replicative longevity than memory cells. The authors of this paper say this might not give the same results if externally stimulated replication was being used, since this can go on for way longer than the capacity for the cells to secrete interleukin-2, which triggers replication under autocrine replication, and that it doesn't necessarily follow that telomere length is the determining cause of senescence. Telomerase activity is upregulated in T cells when stimulated with CD3 and CD28 simultaneously but this might not happen optimally under various experimental setups, and might not happen optimally in-vivo due to decreased expression of CD28 with age. This, they say, might be the driving mechanism to senescence.

From small experiments they ran on oldish (<35 PD) and older (>43 PD) CD4+ cells, they noticed an upregulation of three mitotic inhibitors (p16-INK4alpha, p21-WAF, and p27-kip1) which suggest that upregulation of mitotic inhibitors might be an alternative hypothesis as the cause of senescence.
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Abstract follows:

Replicative senescence may compromise T cell-dependent immune responses to intermittent or chronic antigenic stimulation. While the impact of senescence in vivo remains hard to ascertain, clonal cultures of T cells in vitro provide models for longitudinal studies of aging in well-defined populations. Functional and phenotypic studies as well as investigations into average and maximal longevity of T cells can be performed conveniently with these cloned cells (the former in fact only with cloned cells). Many of the age-associated alterations observed during culture in vitro have also been noted ex vivo in T cells from the elderly.

Moreover, under circumstances where large numbers of antigen- and function-specific T cells may be required, for example for adoptive immunotherapy, the in vitro longevity of the cells may be critically important to successful outcome. These considerations are discussed in the following commentary in the context of immunotherapy of cancer.

The Telomere Shortening Signal May Be Explained by a Fountain Mechanism Modulating the Expression of Eukaryotic Genes

Summary: Speculation on the mechanism involved in telomere-shortening bringing about cell senescence

Interestingness: 4

Paper by AM Olovnikov in the Journal of Anti-Aging Medicine, Volume 2, Issue 1, Spring 1999.

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This dude hypothesised that telomere shortening was the trigger for cell senescence and the existence of telomerase back in the 70s. He now gets to publish whatever he wants about telomeres like the hypothesis in this paper. By the present time (2010) biologists probably know if the theory has any merit but I don't, so to me it's still interesting. The idea is more about basic cell biology than about aging. Its only link with aging is that it explains cell senescence via telomere shortening.

The theory tries to explain how it is that telomere shortening causes senescence. It proposes that some bits of RNA bind to and open Ca2+ and Zn2+ channels on the nuclear membrane, and that the influxes of these ions into the nucleus are critical to the transcription of some/most genes. When telomeres shorten, they would physically pull genes near the telomeres out of the areas where these ion influxes happen and therefore they would stop being transcribed, or at least their transcription patterns would be significantly altered. From what I can tell, the specific bits of RNA, which he calls fountain RNAs (fRNAs), and the importance of the ions to transcription are both speculation.

He says that the location and orientation of the chromosomes between G1 and S phase are nonrandom. The telomeres attach to the nuclear membrane, and the bits of attachment are a reinforced section of the membrane that lack the ion channels in question. As the genes near the telomeres get pulled in closer to the membrane, they would also miss out on the ions.

The fRNAs would be composed of two sections, one that would bind to a section of the genome close to the genes that are going to be induced by the ions, and the other section to the ion channels. The sections of the genome to which the fRNA binds to are called converters. The section of the fRNAs that bind to them would vary depending on which section of the genome the fRNA is meant to stimulate. The other section of the fRNA that binds to the channels, in order to open them, would be constant per channel type, Ca2+ and Zn2+ (although the choice of these two doesn't seem central to the theory, and Mg2+ is listed as another option), but the fRNA wouldn't be able to bind to the channel without having first bound to its converter. The activation of the bits of DNA that code for the fRNAs themselves, called modulators, could themselves be controlled by the ionic fluxes so all sorts of feedback loops and modulation of gene expression would exist.

I had problems distinguishing which bits of the paper were speculation and which parts are presented as evidence. From what I can tell, the following are some of the snippets given as supporting evidence for the theory:

• Ca2+ can increase both transcriptional activity and mRNA stability, increases promoters and RNA levels
• There are Ca2+ releasing channels in the inner nuclear membrane and the nuclear envelope has a store of Ca2+
• Zn2+ involvement in zinc fingers, and their involvement in everything DNA
• Explains the long spacers between genes as spacers decoupling the ionic activation between the genes

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Abstract follows:

We propose a possible mechanism for the telomere shortening signal. The suggested solution of this as yet unsolved enigma—how cell senescence is causally linked to telomere short-ening—is based on a "fountain theory" of modulation of eukaryotic gene expression, in which gene expression is modulated by ionic channels of the inner nuclear membrane. These Ca2+ and Zn2+ channels are opened transiently through the action of a special small nuclear RNA (the fountain RNA or fRNA) on the ionic channels as conformational changes of the fRNA and channel-forming protein occur. Specific Ca2+ and Zn2+ ion channels allow these ions to pass from the perinuclear lumen to the nucleoplasmic gene surroundings. The resultant change of ionic concentration in close vicinity to certain genes, in turn, will alter some in- properties (e.g., mRNA stability, transcript maturation, chromatin configuration, transcriptional activity, and so forth).

Such fRNA-dependent ionic "fountains," may serve as a major mechanism regulating quantitative gene (phenotypic) expression in eukaryotes. We suggest that among metal-activated transcription factors, zinc-finger nuclear proteins evolved, and they are used in the nucleus as an alternative, noncalcium, path of gene-activity modulation, by means of fRNA-dependent channels, increasing the versatility of a fountain system.

We further propose that telomeres are anchored—in a compacted state—to special reinforcing shields, which are parts of the nuclear lamina along the inner nuclear membrane. This may be particularly true between Gl and S phases of the cell cycle, when chromosomes have nonrandom allocation within a nuclear space and telomeres are compacted and serve as "spacers" between the subtelomeric chromosome and the inner nuclear membrane. Each reinforcing shield would cover a portion of the inner nuclear membrane and, in doing so, prohibit the action of fRNA-dependent ion channels, causing an ionic "dead zone" in the nuclear membrane located immediately beneath the shield. When telomeres are long (e.g., in young cells), subtelomeric genes are located at a relatively greater distance from such dead zones; when telomeres shorten and reach the critical threshold, subtelomeric genes become closer to the dead zone and are deprived of contact with active ion channels. Shortening of the telomere—and therefore of the distance of subtelomeric genes from the dead zone—alters subtelomeric gene expression, decreases the functional capabilities of the cell, and results in cell senescence.

In some species, such subtelomeric genes may encode the fRNAs themselves, in addition to structural genes. If modulator genes—coding for fRNAs—require the ion fountains for optimal expression, then other structural genes (in turn modulated by such genes) will inevitably show senescence-associated gene expression as the telomere shortens. Such an alteration of gene expression, and the consequent dysfunction in cellular homeostasis, are typical of senescing cells.