TERT Expression Research Articles

The telomerase RNA component Terc is required for the tumour‐promoting effects of Tert overexpression

A role for the telomerase reverse transcriptase subunit (Tert) in tumorigenesis independent of telomere length is emerging. K5-Tert mice, which overexpress Tert in the skin, show increased tumorigenesis and faster wound healing than wild-type controls. Here, we demonstrate that the telomerase RNA component Terc is necessary to mediate these effects of Tert overexpression. In contrast to K5-Tert mice, K5-Tert mice in a Terc-deficient background, K5-Tert/Terc-/-, do not show increased tumorigenesis or increased wound healing compared with wild-type controls. Indeed, K5-Tert/Terc-/- mice show a reduction in tumour growth compared with Terc-/- controls, indicating an inhibitory effect of Tert overexpression in the absence of Terc. These results indicate that the tumour-promoting effects of Tert overexpression require the formation of Tert-Terc complexes. In addition, we show that the increased expression of Tert in the absence of Terc has an inhibitory effect on tumorigenesis, independently of telomere length and telomerase activity. These findings highlight Terc as a target for telomerase-based anticancer therapies.

Expression of Telomerase Extends the Lifespan and Enhances Osteogenic Differentiation of Adipose Tissue–Derived Stromal Cells

Expression of TERT, the catalytic protein subunit of the telomerase complex, can be used to generate cell lines that expand indefinitely and retain multilineage potential. We have created immortal adipose stromal cell lines (ATSCs) by stably transducing nonhuman primate-derived ATSCs with a retroviral vector expressing TERT. Transduced cells (ATSC-TERT) had an increased level of telomerase activity and increased mean telomere length in the absence of malignant cellular transformation. Long-term culture of the ATSC-TERT cells demonstrated that the cells retain the ability to undergo differentiation along multiple lineages such as adipogenic, chondrogenic, and neurogenic. Untransduced cells demonstrated markedly reduced multilineage and self-renewal potentials after 12 passages in vitro. To determine the functional role of telomerase during osteogenesis, we examined osteogenic differentiation potential of ATSC-TERT cells in vitro. Compared with naive ATSCs, which typically begin to accumulate calcium after 3-4 weeks of induction by osteogenic differentiation medium, ATSC-TERT cells were found to accumulate significant amounts of calcium after only 1 week of culture in osteogenic induction medium. The cells have increased production of osteoblastic markers, such as AP2, osteoblast-specific factor 2, chondroitin sulfate proteoglycan 4, and the tumor necrosis factor receptor superfamily, compared with control ATSCs, indicating that telomerase expression may aid in maintaining the osteogenic stem cell pool during in vitro expansion. These results show that ectopic expression of the telomerase gene in nonhuman primate ATSCs prevents senescence-associated impairment of osteoblast functions and that telomerase therapy may be a useful strategy for bone regeneration and repair.

The Protein Subunit of Telomerase Is Downregulated with Normal Myeloid Differentiation, but Reactivated in Leukemic Progenitors.

Telomeres are comprised of TTAGGG nucleotide repeats and a large protein complex that together protect chromosomal ends. Telomeres shorten progressively with cell division in the absence of telomerase, a reverse transcriptase that synthesizes telomere repeats. Critical telomere shortening has potent effects on cancer development, either blocking cancer progression by impairing proliferation and survival or promoting cancer by enhancing chromosomal instability. Although telomerase is upregulated in the vast majority of human cancers, where it is thought to endow cancer cells with unlimited proliferative capacity, the regulation of telomerase during the course of normal lineage development or during leukemogenesis remains unclear. Here, we show that expression of TERT, the protein catalytic subunit of telomerase, is tightly regulated during myeloid development in both human and mouse. Real time PCR analysis of RNA from purified human stem cell and progenitor cell populations showed that TERT mRNA was highest in human HSCs (100%, values normalized to HSC). TERT mRNA remained high in common myeloid progenitors (CMP, 47%), but was dramatically decreased with differentiation to granulocyte-monocyte progenitors (GMP, 3%) or megakaryocyte-erythroid progenitors (MEP, 0.2%). TERT was similarly down regulated during normal myeloid development in mouse bone marrow. In marked contrast, TERT was reactivated in leukemic GMP in a transgenic Faslpr/lprhMRP8BCL2 mouse model of acute myelogenous leukemia. Notably, transplantation experiments demonstrated that leukemic GMP provided the most robust leukemic transplantation potential compared with HSC or blasts suggesting that TERT reactivation may provide an important final step in leukemic pathogenesis by enhancing the replicative life-span of leukemic progenitors. Currently, we are in the process of assessing TERT regulation in human leukemic progenitors isolated from patients with CML in chronic, accelerated and blast phases. Together, these data indicate that TERT is most highly expressed in stem cells and multipotent progenitors. This pattern of TERT expression may reflect a need for efficient telomere maintenance in these stem/progenitor compartments. Furthermore, these data are consistent with our recent data showing novel, telomere length-independent functions for TERT in stem/progenitor cell compartments.

Mutations in TERT, the Gene Encoding Telomerase Reverse Transcriptase, in “Acquired” Aplastic Anemia Inhibit Enzymatic Function by a Dominant Negative Mechanism of Action.

About one third of patients with acquired aplastic anemia (AA) have very short telomeres of their peripheral blood leukocytes; short telomeres correlate to long disease duration and poor response to immunosuppressive therapy. We have recently shown that some cases of apparently “acquired” AA have mutations in TERC, the gene encoding the RNA component of telomerase (Fogarty et al., Lancet 2003; 632:1628; Yamaguchi et al., Blood 2003; 102:916). In the current study, we examined TERT, the gene encoding the telomerase reverse transcriptase, by sequencing the gene's exons and proximal promoter region in peripheral blood DNA samples from 122 patients with AA and 282 controls. Four novel nonsynonymous mutations among five patients, not present in controls, were discovered; three polymorphisms were identified, two nonsynonymous SNPs and one deletion of a single amino acid. To investigate the functional consequences of the mutations, telomere lengths of leukocytes were assessed by flow cytometric fluorescence in situ hybridization (flow-FISH). All patients carrying TERT mutations had markedly short telomeres compared to age-matched controls, as opposed to other AA patients with polymorphisms, whose telomeres were normal. In one patient's kindred, presence of the TERT mutation in other family members correlated to telomere shortening; non-carriers had normal telomeres. Telomerase function in patients' T cells, activated by phytohemagglutinin and interleukin-2 to increase enzymatic activity, was measured by the telomeric repeat amplification protocol (TRAP). In all mutant AA patients evaluated, cell lysates yielded very low or no telomerase activity, as compared to normal controls. We cloned the AA-related mutations into a TERT expression vector and co-transfected these vectors into VA13 cells (with a TERC-containing vector, as this cell line does not have telomerase activity due to absent TERC and TERT expression). All mutant TERT-containing cell lysates were severely deficient in enzymatic activity. TERT gene expression, as evaluated by Northern blot, was normal in cells transfected with the mutated genes. When vectors containing wild-type TERT and individual TERT mutations were cotransfected, telomerase activity was dramatically reduced, in comparison to transfection of wild-type TERT vector only. These results indicate a dominant negative mechanism of action of TERT mutations as responsible for the absence of telomerase activity in AA patients' cells. Family members lacking telomerase activity have short telomeres but appear physically normal and have no hematological abnormalities. In a provisional model of the telomere repair complex and marrow failure, mutations in DKC1 and the stability regions of TERC cause dyskeratosis congenita, with early presentation and associated physical anomalies. Mutations that affect the enzyme-binding region of TERC and in TERT, the reverse transcriptase itself, lead to a constitutionally reduced stem cell compartment and appear to be genetic risk factors for the development of “acquired” aplastic anemia.

RNA expression of human telomerase subunits TR and TERT is differentially affected by melatonin receptor agonists in the MCF-7 tumor cell line

The RNA expression levels of human catalytic subunit (TERT) and the RNA subunit (TR) of telomerase were analysed after treatment with the agonists of the membrane receptor (S 20098) and the nuclear receptor (CGP 52608) for melatonin in the MCF-7 human breast tumor cell line. Neither membrane nor nuclear signalling affected the RNA steady-state levels of the TR subunit of telomerase. On the contrary, we observed a significant decreased in the RNA levels of TERT after treatment with CGP 52608 while S 20098 produced a significant increase in the RNA levels of TERT. These results support a cross-talk between membrane and nuclear melatonin signalling and provide new data on the hormonal regulation of telomerase function.

Isolation and expression of the reverse transcriptase component of the Canis familiaris telomerase ribonucleoprotein (dogTERT)

The enzyme telomerase plays a crucial role in cellular proliferation and tumorigenesis. Telomerase is an RNA-directed DNA polymerase composed minimally of an RNA subunit (TR) and a catalytic protein component (TERT). The protein component acts as a reverse transcriptase (RT) and catalyses the addition of telomeric repeats onto the ends of chromosomes using the RNA subunit as a template. While both the RNA and catalytic subunits are essential for telomerase activity, the TERT component of telomerase is thought to be the primary determinant for enzyme activity as expression of TERT is largely limited to cells with telomerase activity. We describe here the isolation and sequence characterization of the telomerase catalytic subunit from Canis familiaris (dog), dogTERT. The predicted protein consists of 1123-aa residues and contains all the signature motifs of the TERT family members. Sequence comparisons with previously identified mammalian TERT proteins demonstrate that dogTERT shows the highest level of sequence similarity to the human TERT protein, supporting the dog as a model system for telomerase-based studies. Further, we demonstrate that TERT mRNA expression is associated with telomerase activity in canine-cultured cells, similar to TERT expression in human cells. This data will allow for further investigation of telomerase in canine malignancies as well as the development of the dog as a model system for human telomerase investigations.

Constitutive expression of tert in thymocytes leads to increased incidence and dissemination of T-cell lymphoma in Lck-Tert mice.

Here we describe a new mouse model with constitutive expression of the catalytic subunit of telomerase (Tert) targeted to thymocytes and peripheral T cells (Lck-Tert mice). Two independent Lck-Tert mouse lines showed higher incidences of spontaneous T-cell lymphoma than the corresponding age-matched wild-type controls, indicating that constitutive expression of Tert promotes lymphoma. Interestingly, T-cell lymphomas in Lck-Tert mice were more disseminated than those in wild-type controls and affected both lymphoid and nonlymphoid tissues, while nonlymphoid tissues were never affected with lymphoma in age-matched wild-type controls. Importantly, these roles of Tert constitutive expression in promoting tumor progression and dissemination were independent of the role of telomerase in telomere length maintenance, since telomere length distributions on a single-cell basis were identical in Lck-Tert and wild-type thymocytes. Finally, Tert constitutive expression did not interfere with telomere capping in Lck-Tert primary thymocytes, although it resulted in greater chromosomal instability upon gamma irradiation in Lck-Tert primary lymphocytes than in controls, suggesting that Tert overexpression may interfere with the cellular response to DNA damage.

Telomerase Activity is Constitutively Expressed in the Murine CD8+T Cells and Controlled Transcriptionally and Post-Translationally

Background: Telomerase, a ribonucleoprotein enzyme capable of synthesizing telomeric repeats, attracts attention for its possible role in determining the replicative capacity of normal somatic cells, transformed cells, and cells of the germline lineage. Differently from normal somatic cells with no telomerase activity, normal lymphocytes has been reported to have telomerase activity comparable to that found in transformed cells during development and activation, which substantiate a role in supporting the capacity of lymphocytes for extensive clonal expansion. Methods: Here, in order to define the telomerase regulation in murine T lymphocytes, telomerase activity in cloned murine CD8 + T cells and naive CD8 + T cells isolated from C57BL/6 mice was examined. Next, the regulatory mechanism of telomerase activity at transcriptional and post- translational levels was investigated by determining the expression level of the TERT protein, a key component for telomerase activity. Results: It was demonstrated that telomerase activity was expressed in an inactivated state as well as in an activated state in the murine CD8 + T lymphocytes by using TRAP assay. The increase of telomerase activity was partially dependent on the net increase of TERT expression. Also, telomerase activity was decreased after treatment with protein kinase inhibitors, indicating that telomerase activation was prevented by inhibition of phosphorylation. Conclusion: Therefore, these results suggest that telomerase activity is constitutively expressed in the murine resting T lymphocytes and controlled by both transcriptional regulation and post-translational modifications. (Immune Network 2004;4(3):166-175)

Molecular cloning of the canine telomerase reverse transcriptase gene and its expression in neoplastic and non-neoplastic cells.

To perform molecular cloning of the canine telomerase reverse transcriptase (TERT) gene and determine its expression in neoplastic and non-neoplastic cells. 9 canine tumor cell lines derived from various neoplasms, 16 primary canine tumors, and tissues from 15 normal canine organs. Tumor cell lines were derived from canine tumors that included osteosarcoma, mammary gland adenocarcinoma, melanoma, acute lymphoblastic leukemia, lymphoma, and mastocytoma and a canine primary fibroblast culture. Canine TERT complementary DNA (cDNA) was amplified by use of polymerase chain reaction (PCR) and sequenced. Expression of TERT mRNA was examined by reverse transcription (RT)-PCR assay. Telomerase activity was measured by use of the telomeric repeat amplification protocol assay. The canine TERT cDNA clone was 237 base pairs in length and contained a central region encoding the reverse transcriptase motif 2. Expression of TERT mRNA was detected in canine tumor cell lines that had telomerase activity but not in telomerase-negative canine primary fibroblasts. The TERT mRNA was detected in 13 of 16 canine tumor tissues and several normal tissues such as liver, ovary, lymph node, and thymus. A significant correlation between TERT expression level and telomerase activity was noted. Expression of TERT mRNA was closely associated with telomerase activity in neoplastic cells as well as some non-neoplastic cells from dogs. In addition to telomerase activity, expression of TERT mRNA can be used as a marker of tumor cells.

Karyotype instability and anchorage-independent growth in telomerase-immortalized fibroblasts from two centenarian individuals

Several reports have shown that the ectopic expression of the human telomerase catalytic subunit gene (h TERT) leads to an indefinite extension of the life span of human fibroblasts cultured in vitro without the appearance of cancer-associated changes. We infected two fibroblast strains derived from centenarian individuals with an h TERT containing retrovirus and isolated transduced massive populations (cen2tel and cen3tel). In both populations, h TERT expression reconstituted telomerase activity and extended the life span. In cen2tel, a net telomere lengthening was observed while, in cen3tel, telomeres stabilized at a length lower than that detected in senescent parental cells. Interestingly, both cen2tel and cen3tel cells developed chromosome anomalies, numerical first and structural thereafter. Moreover, cen3tel cells acquired the ability to grow in the absence of solid support, a typical feature of transformed cells. The results we present here highlight an unexpected possible outcome of cellular immortalization driven by telomerase reactivation, and indicate that, in some cases, an artificial extension of cellular replicative capacity can increase the probability of occurrence of genomic alterations, which can lead to cellular transformation.

Loss of telomerase activity may be a potential favorable prognostic marker in lung carcinomas

Many cancer and immortal cells exhibit telomerase activity that stabilizes telomere lengths, possibly contributing to cell immortality and carcinogenesis. The aim of this study was to elucidate the clinicopathological relationship between telomerase activity and telomerase reverse transcriptase subunit (hTERT) status in non small cell lung cancer. hTERT status in non small cell lung cancer using telomeric repeat amplification protocol (TRAP) and RT-PCR assay, respectively. Telomerase activity and hTERT were detected in 85.7 and 80.3% of cancerous tissues, respectively. Telomerase activity does not correlate with clinicopathological variables. However, there was an association between p53-correlated expression and hTERT negative status. Lung cancer patients without telomerase activity survived for a significantly longer period than those with telomerase activity. In addition, hTERT was not associated with the prognosis. TERT expression did not correlate well with any clinical parameter. Reactivated telomerase activity may be a poor prognostic factor in NSCLCs.

Seizures and Tissue Injury Induce Telomerase in Hippocampal Microglial Cells

Telomerase is a reverse transcriptase that adds repeats of a six-base DNA sequence to chromosome ends and thereby prevents their shortening during successive cell divisions. Telomerase activity and expression of the catalytic subunit of telomerase (TERT) are high in brain cells during embryonic development, but are undetectable in the adult. We now report that telomerase activity and expression of TERT are induced in the hippocampus of adult mice after administration of the seizure-inducing excitotoxin kainate. Telomerase activity is present at 24 h and 7 days after kainate administration, but is no longer detectable at 4 weeks. Because the time course of telomerase induction was similar to the time course of microglial activation, we performed studies to determine whether microglia were the source of the telomerase activity. Examination of brain sections immunostained with a TERT antibody and an antibody against a microglia-specific antigen revealed that TERT was not detectable in the uninjured brain, and was present in microglia 24 h and 7 days after kainate administration. This is the first evidence that telomerase can be induced by brain injury and the first evidence that TERT can be expressed in microglia, suggesting roles for telomerase in microglial responses to brain injury.

Properties of a Fetal Multipotent Neural Stem Cell (NEP Cell)

Multipotent neural stem cells (NSCs) present in the developing neural tube (E10.5, neuroepithelial cells; NEP) were examined for the expression of candidate stem cell markers, and the expression of these markers was compared with later appearing precursor cells (E14.5) that can be distinguished by the expression of embryonic neural cell adhesion molecule (E-NCAM) and A2B5. NEP cells possess gap junctions, express connexins, and appear to lack long cilia. Most candidate markers, including Nestin, Presenilin, Notch, and Numb, were expressed by both NEP cells as well as other cell populations. Fibroblast growth factor receptor 4 (FGFR4), Frizzled 9 (Fz9), and SRY box-containing gene 2 (Sox2) as assessed by immunocytochemistry and in situ hybridization are markers that appear to distinguish NSCs from other precursor cells. Neither Hoechst 33342 nor rhodamine-123 staining, telomerase (Tert) expression, telomerase activity, or breakpoint cluster region protein 1 (Bcrp1) transporter expression could be used to distinguish NEP stem cells from other dividing cells. NEP cells, however, lacked expression of several lineage markers that are expressed by later appearing cells. These included absence of expression of CD44, E-NCAM, A2B5, epidermal growth factor receptor (EGFR), and platelet-derived growth factor receptor-alpha (PDGFRα), suggesting that negative selection using cell surface epitopes could be used to isolate stem cell populations from mixed cultures of cells. Using mixed cultures of cells isolated from E14.5 stage embryos, we show that NEP cells can be enriched by depleting differentiating cells that express E-NCAM or A2B5 immunoreactivity. Overall, our results show that a spectrum of markers used in combination can reliably distinguish multipotent NSCs from other precursor cells as well as differentiated cells present in the CNS.

Properties of a Fetal Multipotent Neural Stem Cell (NEP Cell)

Multipotent neural stem cells (NSCs) present in the developing neural tube (E10.5, neuroepithelial cells; NEP) were examined for the expression of candidate stem cell markers, and the expression of these markers was compared with later appearing precursor cells (E14.5) that can be distinguished by the expression of embryonic neural cell adhesion molecule (E-NCAM) and A2B5. NEP cells possess gap junctions, express connexins, and appear to lack long cilia. Most candidate markers, including Nestin, Presenilin, Notch, and Numb, were expressed by both NEP cells as well as other cell populations. Fibroblast growth factor receptor 4 (FGFR4), Frizzled 9 (Fz9), and SRY box-containing gene 2 (Sox2) as assessed by immunocytochemistry and in situ hybridization are markers that appear to distinguish NSCs from other precursor cells. Neither Hoechst 33342 nor rhodamine-123 staining, telomerase (Tert) expression, telomerase activity, or breakpoint cluster region protein 1 (Bcrp1) transporter expression could be used to distinguish NEP stem cells from other dividing cells. NEP cells, however, lacked expression of several lineage markers that are expressed by later appearing cells. These included absence of expression of CD44, E-NCAM, A2B5, epidermal growth factor receptor (EGFR), and platelet-derived growth factor receptor-alpha (PDGFRα), suggesting that negative selection using cell surface epitopes could be used to isolate stem cell populations from mixed cultures of cells. Using mixed cultures of cells isolated from E14.5 stage embryos, we show that NEP cells can be enriched by depleting differentiating cells that express E-NCAM or A2B5 immunoreactivity. Overall, our results show that a spectrum of markers used in combination can reliably distinguish multipotent NSCs from other precursor cells as well as differentiated cells present in the CNS.

Variant forms of upstream stimulatory factors (USFs) control the promoter activity of hTERT, the human gene encoding the catalytic subunit of telomerase

It is known that Myc regulates the expression of TERT, the telomerase catalytic subunit gene, by binding to E box. Here we show that another E box-binding protein, upstream stimulatory factor (USF), also regulates TERT expression. Specifically, the N-terminally truncated form of USF2 is present in telomerase-negative/resting human lymphocytes, but not in telomerase-positive/phytohemagglutinin-activated lymphocytes. In electrophoretic mobility shift assay, both full-length and truncated USF2s bound to the TERT E box. In a transient expression assay, the truncated USF had a dominant-negative effect on both exogenous full-length USF and endogenous positive regulators for activating TERT expression. These results suggest that the differential abundance of truncated USF2 may regulate telomerase activity during lymphocyte activation.

Cooperation of hTERT, SV40 T Antigen and Oncogenic Ras in Tumorigenesis: A Cell Transplantation Model Using Bovine Adrenocortical Cells

Expression of TERT, the reverse transcriptase component of telomerase, is necessary to convert normal human cells to cancer cells. Despite this, “telomerization” by hTERT does not appear to alter the normal properties of cells. In a cell transplantation model in which bovine adrenocortical cells form vascularized tissue structures beneath the kidney capsule in scid mice, telomerization does not perturb the functional tissue-forming capacity of the cells. This cell transplantation model was used to study the cooperation of hTERT with SV40 T antigen (SV40 TAg) and oncogenic Ras in tumorigenesis. Only cells expressing all three genes were tumorigenic; this required large T, but not small t, antigen. These cells produced a continuously expanding tissue mass; they were invasive with respect to adjacent organs and eventually destroyed the kidney. Cells expressing only hTERT or only Ras produced minimally altered tissues. In contrast, SV40 TAg alone produced noninvasive nodules beneath the kidney capsule that had high proliferation rates balanced by high rates of apoptosis. The use of cell transplantation techniques in a cell type that is able to form tissue structures with or without full neoplastic conversion allows the phenotypes produced by individual cooperating oncogenes to be observed.

Telomerase activity in primary cultures of normal adrenocortical cells.

Telomerase activity was measured in isolated cells from bovine and human adrenal cortex, in cells in primary culture, in cells in later passages in culture, and in cells genetically modified by expression of hTERT (human telomerase reverse transcriptase). Telomerase activity in freshly isolated bovine adrenocortical cells and in human adrenal cells from donors of various ages (6-79 years) was very low or undetected. However, primary bovine adrenocortical cell cultures were strongly positive for telomerase activity, and primary human adrenocortical cell cultures were weakly positive. Both cell types proliferate in primary culture but proliferation of bovine cells is much more vigorous. When primary bovine cells were subcultured to make successively secondary and tertiary cultures, telomerase activity declined strongly, and was undetected by the third passage. There was only a slight decrease in growth rate over this period. Levels of the telomerase RNA component did not change with passage number when assessed by semi-quantitative competitive RT-PCR. When both bovine and human cells were infected with a retrovirus encoding hTERT, telomerase activity in the cells was very high. We conclude that in the adrenal cortex, as in some other tissues, TERT expression is regulated and upregulation of telomerase activity is associated with rapid proliferation in primary culture. Telomerase activity is not maintained, and introduction of TERT is required for stable telomerase activity and for immortalization.

In Situ Detection of hTERT mRNA Relates to Ki-67 Labeling Index in Papillary Thyroid Carcinoma

Background. Telomerase is activated in most human cancers but is inactivate in adult somatic tissues except for some proliferating cell lineages. The maintenance of telomerase activity may be a critical step of cellular immortalization and transformation.Materials and methods. We analyzed the expression of human telomerase reverse transcriptase (hTERT) using in situ hybridization and compared it to Ki-67 immunoreactivity in 29 cases of papillary thyroid carcinoma (PTC) and 17 cases of benign thyroid disease.Results. The hTERT messenger RNA (mRNA) was expressed in the cytoplasm of carcinoma cells with moderate (n = 10) to strong intensity (n = 10) in 69% (20 of 29) PTC cases. Human TERT was found in only 29% (5 of 17) cases of benign thyroid disease. Human TERT gene expression was preferentially detected in PTC (P = 0.021). The Ki-67 labeling index was observed in 16 cases of PTC (16 of 29; 55.2%). This result was significantly different from that of benign thyroid disease (P = 0.014). The Ki-67 labeling index related to the intensity of hTERT mRNA expression (r = 0.51; P = 0.005) and was inversely associated with the follicular variant of PTC (r = −0.413; P = 0.026). No statistically significant difference was found between hTERT expression and histological subtype of PTC.Conclusions. Our results demonstrated that expression of hTERT could be detected using in situ hybridization in PTCs and was significantly distinguishable from that of benign thyroid disease. Human TERT expression was related to the Ki-67 labeling index, indicating that coupling of telomerase activation with cell proliferation was the associated mechanism for tumorigenesis.

Differential regulation of telomerase activity and TERT expression during brain development in mice

Differential regulation of telomerase activity and TERT expression during brain development in mice

Differential regulation of telomerase activity and TERT expression during brain development in mice.

Telomerase is an enzyme activity consisting of a reverse transcriptase called TERT and an RNA component that adds repeats of a DNA sequence (TTAGGG) to the ends of chromosomes, thereby preventing their shortening. Associations between telomerase activity and proliferation and differentiation of neural tumor cells and neural stem cells have been reported, but the role of telomerase in brain development is unknown. We now report analyses of telomerase activity, TERT mRNA levels and levels of mRNAs encoding the telomere-associated proteins TRF1 and TRF2 in three different brain regions (brainstem, hippocampus and cerebral cortex) and the eye of mice at increasing developmental time points. Telomerase activity is high in the brain at embryonic day 13 (E13), declines markedly between E13 and E18, remains at a low level until postnatal day 3 (P3) and becomes undetectable by P10. Surprisingly, the temporal pattern of change in telomerase activity is not paralleled by a decrease in levels of TERT mRNA that remain elevated from E13 to P5 (with fluctuations during this time window that vary among brain regions), and then decrease to a lower level that is maintained into adulthood. TRF1 and TRF2 mRNA levels are relatively constant throughout brain development. Our data are consistent with a role for telomerase activity in proliferation of neural progenitor cells, and further suggest that TERT may play roles in neuronal differentiation and survival. The dissociation between TERT expression and telomerase activity is a novel finding that suggests biological functions for TERT in addition to telomere maintenance. J. Neurosci. Res. 64:252-260, 2001. Published 2001 Wiley-Liss, Inc.