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Contents
1. Telomere biology . . . . . . . 1.1. Introduction . . . . . . . 1.2. End-replication problem 1.3. Telomere hypothesis . . . 1.4. Telomere conguration . 1.5. Telomerase holoenzyme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 30 30 30 30 31 32 32 33 33 33 34 35 35 36 36 36 37 37 37 37 38 38 38 38 39 40
2. Aging . . . . . . . . . . . . . . . . . . . . . . 2.1. Senescence and telomerase . . . . . . . 2.2. Genetic disorders and telomeres . . . . 2.3. Hematopoietic system and telomerase. 2.3.1. Stem cells . . . . . . . . . . . . . 2.3.2. Peripheral blood leukocytes . . .
3. Cancer and telomerase . . . . . . . . . . . . . . . . . . 3.1. Survey of telomerase and cancer. . . . . . . . . . 3.2. Role of telomerase in malignant transformation . 3.3. Methods of telomerase acquisition. . . . . . . . . 3.4. Prognostic implications of telomerase detection . 3.5. Residual disease . . . . . . . . . . . . . . . . . . . 3.6. Diagnostic potential . . . . . . . . . . . . . . . . . 3.7. Therapeutic potential . . . . . . . . . . . . . . . . 3.7.1. Telomerase inhibitors . . . . . . . . . . . . 3.7.2. Immunotherapy and telomerase . . . . . . 3.7.3. Chemoprevention and telomerase . . . . .
* Corresponding author. Tel.: + 1-214-648-3282; fax: + 1-214-648-8694. E -mail addresses: meaghan.granger@utsouthwestern.edu (M.P. Granger), jerry.shay@utsouthwestern.edu (J.W. Shay).
woodring.wright@utsouthwestern.edu
(W.E.
Wright),
1040-8428/02/$ - see front matter 2002 Elsevier Science Ireland Ltd. All rights reserved. PII: S 1 0 4 0 - 8 4 2 8 ( 0 1 ) 0 0 1 8 8 - 3
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Abstract The telomeretelomerase hypothesis is the science of cellular aging (senescence) and cancer. The ends of chromosomes, telomeres, count the number of divisions a cell can undergo before entering permanent growth arrest. As divisions are being counted, events occur on the cellular and molecular level, which may either delay or hasten this arrest. As humans age, a particular concern is the accumulation of events that lead to the progression of cancer. Telomerase is a mechanism that most normal cells do not possess, but almost all cancer cells acquire, to overcome their mortality and extend their lifespan. This review aims to provide a comprehensive understanding of the role of telomerase in cancer development, progression, diagnosis, and in the future, treatment. The ultimate goal of telomerase research is to use our understanding to develop anti-telomerase therapies, an almost universal tumor target. 2002 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: Telomere; Telomerase; Senescence; Replicative aging; Cancer; Immunosenescence; Telomerase inhibitors
1. Telomere biology
1.1. Introduction
After fertilization and mixing of 23 paternal and 23 maternal chromosomes, human life begins as a single cell with 46 chromosomes whose initial function is to divide. Each new generation of daughter cells successively divides until it forms and develops into a complex, differentiated organism. With each division, the genetic code is transferred as our chromosomes are replicated and distributed into the daughter cells. There are many cellular mechanisms in place to ensure that the transfer of information is done in a reliable, accurate, and efcient manner throughout the many duplications required over a human lifetime. Two of the mechanisms central to the subject of this review are the semi-conservative replication of DNA and cellular senescence.
end of the chromosome. Thus, the extreme end of the chromosome is not replicated and the telomeres progressively shorten. This is known as the end-replication problem. Fortunately, this problem does not result in the loss of essential genes in that each of the 46 human chromosomes is capped with long repeats of expendable noncoding DNA bases called telomeres (Fig. 1). Loss of the telomeric DNA continues with successive divisions until the telomeres reach such a critically short length that replication is halted. Human cells are estimated to have the potential to undergo on average 60 70 divisions, and at this point the cells growth arrest and enter senescence [1].
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Fig. 1. Metaphase spread of human broblasts visualized in a uorescence microscope. Fluorescence in situ hybridization (FISH) using telomeric probes reveal the red/pink dots at the ends of each chromosome. Each dot identies a telomere and shows the two telomeres per chromosome with a total of 96 telomeres per normal human cell.
now know they are a site of dynamic activity beyond being the biologic timepiece [6]. They have a unique T-looped conguration where the telomere bends back on itself [7]. The overhanging guanine-rich single strand is tucked into the double stranded telomere. This creates a second smaller d-loop by displacing one of the telomere strands. This structure appears to protect the telomeres from end to end fusion with other chromosomes and from cell cycle checkpoints that would otherwise recognize the telomeres as chromosome breaks requiring repair (reviewed in [8]). Proteins that localize specically to telomeric DNA are the duplex telomere binding proteins, TRF1 and TRF2. TRF1 and 2 and their associated proteins have the primary responsibility of stabilizing the complex and forming the t-loop. Some degree of stabilization is intrinsic to the telomere overhang due to the G-rich nature of the TTAGGG repeats that form quadruplex structures. TRF1 is important in intratelomeric coiling [7]. TRF2 also binds along the length of the telomere but appears to be particularly abundant at the base of the t-loop and is important for its stabilization and formation [7]. Their cooperation is similar to two hands tying a knot, the rst hand (TRF1) forms a loop and the second hand (TRF2) tightens the strand and secures it. These duplex telomere DNA binding proteins also have their own associated proteins [9]. Human rap1p is integrated into the t-loop complex and interacts with TRF2, but its specic role in humans is unknown [10].
Tankyrase has the ability to inhibit TRF1, thereby releasing it from the t-complex and allowing telomerase and other enzymes to bind. TIN2 promotes TRF1 function and causes it to bind to the telomere [9]. The DNA damage response complex RAD50/MRE11/ NBS1 also cooperates with TRF2. The MRE11 complex functions conventionally in homologous recombination to repair DNA double strand breaks [11]. At the telomere, however, it is thought to stabilize the d-loop where the single stranded tail invades the duplex telomere. Based on its function in vitro, the role of NBS1 during the S phase may be to unwind the t-loop via a helicase [12].
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Fig. 2. Telomerase holoenzyme. The telomerase holoenzyme adds telomeric repeats, TTAGGG, in two steps (1) elongation and (2) translocation in succession. The enzyme is composed of two primary parts: hTR is the telomerase functional or template RNA portion, and hTERT is the telomerase reverse transcriptase enzymatic portion. The telomeric end can binds to the template region of hTR and is elongated by the addition of the bases complementary to the template via the catalytic subunit (hTERT). The complex then pauses and translocates and repeats the elongation of the telomere (e.g. the human telomerase complex is processive).
combining recombinant foldosome proteins, hTR, and hTERT is sufcient to reconstitute the holoenzyme [16]. The telomerase gene was recently mapped to 5p15.33 as one of the most distal genes on chromosome 5p. This has raised questions about whether its proximity to the telomere might result in it being regulated by telomere position effect mechanisms [17 19]. The introduction of the catalytic protein (hTERT) component of telomerase into normal broblasts and epithelial cells prevents shortening of the telomeres, and results in immortalization [20]. The key role of telomerase in immortalization is to maintain telomere length, not to produce a net increase in length [15]. Transient expression of a cre -excisable telomerase results in a preferential lengthening of the shortest telomeres and an increase in lifespan proportional to the length of the shortest telomere [21]. Likewise, the inhibition of telomerase in immortalized human cells leads to progressive telomere shortening and cell death [22].
2. Aging
Normal cells have a nite number of divisions they can undergo before entering retirement or replicative senescence. Cells removed from older individuals, in general, divide fewer times in culture when compared to cells obtained from younger patients. Replicative senescence is the process by which cells stop dividing due to a genetically programmed event. Normal cells reach a period of growth arrest termed M1, or mortality stage 1, that is controlled by cell cycle regulatory genes p53/p21 and perhaps p16/Rb. There is speculation that M1 might be initiated by the presence of at least one sufciently short telomere and activation of the DNA damage response, although at this growth point most of the 92 telomeres still have several kilobase pairs of telomeric repeats. Other possibilities include regulation by subtelomeric genes or by transcription factors associated with the telomere [9]. If p53 function is altered or blocked (as with SV40 T antigen or E6/E7 papillomavirus proteins) cells continue to divide with progressive telomere shortening until they reach a second stage known as M2, mortality stage 2. It has been established that telomere shortening controls both M1 and M2 [15,24]. The M2 stage is often referred to as crisis at the point where many telomeres have been critically shortened and can no longer protect the telomeres so that chromosome fusion and breakage cycles occur and the cells eventually undergo apoptosis. In human broblasts in vitro that express viral oncogenes, a small number of cells (1 10 7), are able to escape M2 crisis and immortalize by the acquisition of a method for maintaining stable telomeres. This is accomplished through a reactivation oftelomerase in most cells, but alternative lengthening of telomere mechanisms (ALT) exist that use recombination and copy switching to move DNA from one telomere to another [25]. It is believed that replicative senescence decreases the number of mutations that can occur bylimiting the number of times the cell can divide. Properties of senescence are dependent on the number of cell divisions not time. It entails cells entering an irreversible state incapable of proliferation and with altered function. Cells become growth arrested in G1 and are unable to replicate their DNA [26]. What is the relationship between senescence, aging, cancer, and telomerase? Telomeres shorten in aging cell populations in vitro and in vivo (Fig. 3). Human broblasts from fetal tissues can typically undergo 60 80 population doublings (PDs), whereas young adult cells achieve only 2040 doublings, and older adult cells 1020 doublings before entering senescence. It is important to understand the molecular mechanism regulating senescence in oncology because it is the very cellular outcome we are seeking, for the cancer cells to stop dividing [26].
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Fig. 3. Telomere hypothesis. (a) With progressive cell divisions, telomeres shorten until they reach a critical shortened length. At this point, they undergo growth arrest or apoptosis (depending on whether other cellular pathways have been altered) unless they are able to maintain their telomeres to allow for subsequent divisions. (b) Stem cells exhibit a slower rate of telomere shortening because of the intrinsic presence of telomerase in these cells indicating that they may undergo more doubling prior to becoming senescent. (c) Committed peripheral blood lymphocytes (PBL, dotted line) are derived from the stem cell compartment and have a telomere length correlating with their age at the time of commitment. PBLs, upon activation, can have a brief period of telomerase upregulation followed by continued telomeric shortening. (d) Stem cell transplant recipients have accelerated telomeric shortening following transplant and then continued shortening at a rate proportional to the donor stem cells. (e) Cancer cells (dashed lines) may develop at any point in normal and hematopoietic cells and, in most cases, have utilized telomerase to maintain their telomeres. (f) All cells have higher rates of telomere loss from birth to 1 year, somewhat less from 1 to 4 years, followed by consistent loss of 50 100 bp/division.
2.3. Hematopoietic system and telomerase 2.3.1. Stem cells Telomerase activity can be detected in both hematopoietic stem cells and in stem cell populations in other tissues such as skin, hair follicles, small intestine crypt cells, and lymphoid cells. Though the hematopoietic cells possess telomerase, they still have telomeres that shorten. Stem cells that are CD34 +/CD38 have shorter telomeres in adults than the same cell type in fetal and newborn tissue [31]. It is believed that the expression of telomerase in stem cells may help slow down, but does not completely prevent telomere attrition in cells that have a high rate of turnover (Fig. 3). Telomerase activity ensures that the stem cell compartment will be able to handle potentially large expansion demands, preserving the ability to maintain and repair the tissues. Though the telomeres still shorten, the time to critically shortened length may be delayed by telomerase [32]. Studies in stem cell transplant patients have shown that stem cells are on average 0.4 kb shorter in the reconstituted recipient when compared concurrently with the donor (Fig. 3). It is likely that the proliferation demands required to reconstitute the entire hematopoietic system
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results in aging of the cells prematurely by : 15 years [33]. Further, this shortening could eventually be sufcient to contribute to genetic instability and account for some of the secondary neoplasms seen in stem cell transplant patients beyond those attributed to alkylating agents and etoposide [31]. It is unknown whether the cumulative dose of stem cells given patients have an effect on this aging [33]. Telomere lengths in aged hematopoietic stem cells have not been shown to reach a critically shortened length leading to complete senescence. However, the cellularity of the bone marrow compartment is reduced by one-third at the age of 70 years [34]. It has been suggested that the replicative stress of shortening telomeres, particularly in lymphocytes, seen in early childhood and in the elderly might be responsible for the coinciding with the bimodal distribution of some hematopoietic disorders [34]. Acute lymphoblastic leukemia peaks in occurrence in children at an average age of 4 years [35] and again in adults after the age of 45 years [36]. The fact that this disorder is extremely rare during the critical years of childbearing and rearing and more common in early childhood and the elderly may be evidence of an evolutionary tumor protective mechanism.
2.3.2. Peripheral blood leukocytes The aging immune system involves a complex change in the entire system, both constitutionally and functionally. It is clinically apparent that aging individuals are at increased risk for infection, cancer, decreased immunity from previous vaccination, and reactivation of latent disease such as varicella. An overview of the global nature of these changes has recently appeared [37]. T cells, in general, shift from na ve to mature memory types with an increased proportion found in the bone marrow rather than peripheral blood. There are proportionally more CD8 + T-cells than CD4 + . B cells also show increased levels in the bone marrow with overall qualitative defects in antibody production. This is presumed to be from increased somatic mutations affecting Ig-gene rearrangements but is also inuenced by the shift in the T-helper cell population from Th1 to Th2. In contrast to decreased circulation of T and B cells, NK cells are found in increased numbers [37]. Granulocytes show decreased phagocytosis and respiratory burst in aging individuals. Monocytes are more activated, dendritic cells are unchanged, and macrophages increase their production of cytokines. Erythrocytes exhibit a shift in proportions of young to old populations [37]. As in the stem cell compartment, both circulating T and B cells have progressive telomere shortening with age and express low levels of telomerase activity at
rest, but levels transiently increase with stimulation by mitogens (Fig. 3, committed PBL). Interestingly, hTERT (the mRNA component of telomerase) appears to be constant among all lymphocyte stages independent of the level of telomerase activity [38,39]. This is consistent with most normal telomerase positive somatic cell types that still exhibit shortening in spite of the presence of telomerase and could reect alternatively spliced variants of hTERT that are inactive. Several studies have shown that telomere shortening with aging in peripheral blood leukocytes, both T lymphocytes and neutrophils, occurs in at least two phases. First, there is a rapid shortening from birth to 4 years at about 1 kb per year. Next there is a gradual shortening until : 40-years-old of 2050 bp per year and more slowly thereafter (Fig. 3) [34,40]. These reect the complexities due to the presence of telomerase, which may make telomere lengthening and shortening a more dynamic system depending on the hematopoietic requirements of the body. Certainly, the demand for clonal proliferation of a committed lymphocyte may increase or decrease telomere length, but also the telomere length of the originating stem cell can play a role [34]. Replicative senescence is intact in normal T-cells just as in broblasts and other cells. However, the implications of senescence in the immune system are more signicant. Mature T cells are required to give rise to clonal proliferations of cells to respond to foreign antigens upon activation. This cannot occur if the T cells are senescent. Senescence in culture reliably occurs in T lymphocytes, both CD4 + and CD8 + , after 2540 PDs. Thus, each mature T cell is capable of producing : 240 cells, or 1 1012 cells, before senescing [41]. There are signicant functional changes in senescent T cells. The most important being the lack of expression of CD28, which plays a key role in the transduction of IL-2 transcription and receptor expression, cooperation with B cells for antibody production, T cell homing, and signaling the induction of telomerase activity [41]. CD28 is present on 99% of neonatal T cell compared to only 45% of centenarian T cells. Telomeres of CD28 cells are shorter than CD28 + telomeres [41]. The CD28 cells are primarily of the CD8 + subset, which play a pivotal role in cytotoxic functions against cells with endogenously expressed antigens such as virally infected cells and tumor cells. Senescent T cells also acquire resistant to apoptosis that results from an increase in bcl-2 [41]. Telomeric changes in B cells are quite different from T cells. Rather than the steady but slow decline in telomeric length of aging T cells, activated B cells in germinal centers of tonsillar tissue show an increase in
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telomere length from their na ve state. The length then begins to decline once the B cells enter the memory compartment. Telomerase activity is highest in tonsillar B cell germinal centers, which corresponds to the point of longest telomeres. This is possibly a mechanism to protect the telomeres of highly specic B cells from the replicative stress placed on B memory cells [42].
3. Cancer and telomerase Telomerase expression is a hallmark of cancer. Nearly the complete spectrum of human tumors has been shown to be telomerase positive (Fig. 4). In general, malignant tumors are characterized by telomerase expression, indicating the capacity for unlimited proliferation and thus immortality. Most benign tumors are characterized by the absence of telomerase, indicating their limited proliferative capacity, and ultimate senescence.
Fig. 4. Summary of telomerase activity expression in human cancers from a review of the literature. Tumor samples were assayed by the TRAP assay. Percentages in parentheses refer to the number of samples that were telomerase positive compared to matched control tissue. Adapted from Shay and Bacchetti, 1997. Please refer to original article for details and discussion [43]
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over the course of the disease. This increase is accompanied by a net loss in telomeric length [32]. A series of 58 patients showed that high telomerase activity and shorter telomeres had an adverse prognosis [44]. Chronic myeloid leukemia does not show an increase in telomerase activity over peripheral mononuclear cells, however, a shorter TRF length correlates with shorter time to blast crisis phase. Small studies in acute lymphoblastic leukemia have found telomerase activity to be variable [32]. Acute myelogenous leukemia, multiple myeloma, plasma cells leukemia, and nonHodgkins lymphoma all exhibit telomerase positivity [45]. However, Hodgkins lymphoma does not exhibit telomerase activity [46].
are based on the idea that cancer arises by clonal expansion of proliferating cells, and it is the stem cells of epithelial tissues that constitute the pool of proliferating cells. Alternatively, it might be expected that cancer would arise in differentiated cells rather than stem cells since the mass of most tissues is comprised of differentiated cells. Following a mutation that initiates clonal expansion, the pre-malignant cell accumulates other critical mutations such as p53 resulting in genomic instability and continued cell division and further shortening of telomeres. This repetitive, clonal expansion leads to the acquisition of other mutations, loss of heterozygosity and the ultimate upregulation or reactivation of telomerase. This upregulation or reactivation of telomerase permits the stabilization of the telomeres and an immortal state [52].
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cious cytogenetic abnormalities in breast cancer correlate with increased telomerase activity, namely 3q + (site of hTR), 8q + (c-myc), and 17p (p53) [57]. One hundred patients with colorectal cancer were followed for 3 years after surgery. High telomerase activity was found in 44/100 patients at the time of biopsy and correlated with a signicantly (P = 0.01) decreased survival, 81% vs. 43% [58]. A retrospective study in patients with meningioma appeared to predict relapse. In 25 patients that were examined, ve patients had detectable telomerase activity and subsequently relapsed. Twenty-ve patients had no detectable telomerase activity and did not relapse [59]. Glioblastoma is one of the few examples where no correlation has been seen between grade and telomerase activity. TRAP levels have been found to be highly variable both within the same patient and within a series of glioblastomas [60]. These observations of the prognostic utility of telomerase assays have not yet reached the clinic in terms of predicting outcome for patients.
with telomerase activity were ultimately diagnosed as carcinoma and 6/7 without telomerase activity were ultimately diagnosed as benign lesions with a P = 0.0007. The TRAP assay thus has the potential to augment the FNA screening tool in combination with cytology in the early diagnosis of breast cancer [61]. Telomerase activity has all the desired characteristics to be used as a potential cancer-screening tool. It requires a small amount of tissue, can be done on a variety of tissue types or body uids requiring minimal invasiveness, has a sensitive assay, is specic to the malignant state in most instances, and can be done at a low cost [62].
3.7.1. Telomerase inhibitors The RNA template of the telomerase holoenzyme is a popular target for inhibition research using antisense oligonucleotides that are complementary to this region
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of hTR. Regardless of the conguration of telomerase, the template region of hTR must be accessible to bind to the telomeric repeats, which exposes it to inhibition by antisense approaches. The major challenge for this class of drugs is access and stability how to get the oligonucleotides into the cell and then to the enzyme without being degraded by nucleases. One strategy has been to modify the DNA using sugar-modied RNA, such as 2%-O -methyl RNA and the 2%-methoxyethoxy RNA [63]. In the laboratory, telomerase can be inhibited by the introduction of a dominant-negative hTERT gene into the cell. The gene encodes a point-mutated reverse transcriptase crippled hTERT that inhibits wildtype hTERT both by sequestering the available hTR and by competing with the wild-type hTERT for access to the telomeres. In vitro studies have shown that the introduction of the dominant-negative (DNhTERT) into cancer cells inhibits telomerase and leads to progressive telomere shortening and cell death [64]. Wild-type hTERT, DN-hTERT, and control vectors were introduced into 36 M ovarian carcinoma cell lines in culture. After several PDs, the cells were introduced into nude mice to assess for tumorigenicity. The wild-type and control vector cells produced tumors but the DN-hTERT cells did not. The application of this design may be more feasible as the area of gene therapy progresses. Attention has also been given to the reverse transcriptase inhibitor class of drugs, such as AZT, that have been effective in HIV treatment. Unfortunately, it has not been shown to date that this class of agents promotes shortening of telomeres and senescence or apoptosis of the treated cells [63].
agent. Human mammary epithelial cells from women with Li-Fraumeni syndrome are characterized by a mutation in the p53 tumor suppressor gene that makes it nonfunctional. These cells spontaneously immortalize in culture at a reliable frequency. Using a variety of telomerase inhibitors, such as the 2-O -methyl-RNA antisense oligonuclotide, the dominant negative hTERT, or nontoxic concentrations of other chemotherapeutic agents, the rate of in vitro immortalization was signicantly reduced [67]. Other patients at high risk for spontaneous immortalization could benet from this strategy of chemoprevention including those at high risk for lung cancer from smoking or chemical exposure, patients treated for a primary malignancy with a high probability of recurrence, and those with conditions considered premalignant with a high probability of progression.
4. Conclusion A hypothesis gaining support is that the function of cellular senescence is to restrict the number of mutations that can be accumulated by a pre-malignant cell. If one accepts this hypothesis, then counting cell divisions becomes the distinguishing feature of replicative aging. Determining whether replicative aging has relevance to organismal aging remains a fundamental unresolved issue. However, there is mounting experimental support that restoring mortality by inhibiting telomerase in tumors may be an effective therapy and is an area where great progress is anticipated in the near future. Telomere biology is clearly important in replicative aging and cancer. Cancer cells need a mechanism to maintain telomeres, if they are going to divide indenitely, and telomerase solves this problem. The key is to understand how the telomerase holoenzyme and telomere-complex interact to maintain telomere length. The challenge is to learn how to intervene in these processes and exploit our increasing knowledge of telomere biology for cell and tissue engineering as well as the diagnosis and treatment of malignancies.
3.7.2. Immunotherapy and telomerase Recently, Vonderheide, et al. identied a tumor-associated antigen (TAA) that correlates with hTERT expression in an HLA subset of patients. He generated cytotoxic T lymphocytes and demonstrated hTERT specic cytolysis in many tumor lines that spared telomerase positive peripheral blood CD34 + cells. However, since CD40 + activated B cells were lysed, it is possible that the immune system will not function optimally in a clinical setting if it is forced to rely solely on the interaction of antigen processing cells with cytotoxic T cells without activated B cells in the germinal centers [65]. Other investigations have shown similar results with other hTERT peptides that are able to generate a cytotoxic response against tumor cells but not telomerase-positive CD34 + stem cells [66]. 3.7.3. Chemopre6ention and telomerase Telomerase antisense inhibitors have been recently shown to have potential value as a chemopreventative
Reviewers Joachim Lingner, PhD, Swiss Institute for Experimental Cancer Research (ISREC), 155, ch. des Boveresses, CH-1066 Epalinges, Switzerland. Petra Boukamp, PhD, Deutsches Krebsforschungszentrum (DKFZ), Abteilung B0600/FS2, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany. Dr. Go ran Roos, Department of Pathology, Umea University, S-90187 Umea, Sweden.
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Biographies Meaghan P. Granger, MD is a clinical fellow in pediatric Hematology and Oncology at UT Southwestern Medical Center. She received her MD from the University of Arkansas and completed her residency in pediatrics at Vanderbilt University. She is currently conducting telomerase research in the laboratory of Jerry W. Shay, PhD and Woodring E. Wright, MD/ PhD in the Department of Cell Biology. Woodring E. Wright received his BA from Harvard College and then completed his MD/PhD at Stanford University in California, where earned his PhD in the laboratory of Leonard Hayick. He pursued postdoctoral studies at the Pasteur Institute in Paris with Franc ois Gros and then joined the faculty of Southwestern Medical Center where he is currently a professor of Cell Biology. Jerry W. Shay earned his BA and MA at the University of Texas at Austin and his PhD at the University of Kansas at Lawrence. He did his postdoctoral work at the University of Colorado in Boulder with Keith Porter and David Prescott before moving to Dallas where he is currently a professor of Cell Biology at the University of Texas Southwestern Medical Center in Dallas and an Ellison Medical Foundation Senior Scholar. In 1985, Shay and Wright began what has become a very close and productive collaboration. This led to the development of the two-stage model for cellular senescence for which they shared the Allied Signal Award for research on aging in 1995 and in 2001 the American Aging Association Hayick Award. They are both members of Gerons scientic advisory board and have over 15 patents allowed on their telomere and telomerase-based research. Both have served on the Scientic Research Board of the American Foundation for Aging Research.