JOURNAL OF PATHOLOGY, VOL.

180: 233-235 (1996)

REVIEW

TELOMERES AND DISEASE

Telomeres define the natural ends of linear chromo- that contains the C-rich strand of one and a half
somes and their sequence organization is highly con- telomere repeats in the ‘template r e g i ~ n ’ . ~The
J ~ tem-
served in the majority of (though not all) eukaryotes. plate region anneals to the last few bases of a chromo-
Broken chromosomes that lack telomeres can fuse to some and the remaining bases in the template region are
one another. The fused dicentric chromosome enters a reverse-transcribed by the protein components, resulting
cycle of anaphase bridge formation, further uncon- in the addition of a new telomere repeat onto the G-rich
trolled chromosome breakage, and fusion, usually strand. Furthermore, the processive nature of the human
resulting in cell death.’ Telomeres therefore ‘cap’ or telomerase activity indicates that the enzyme ‘translo-
protect the chromosome from degradation and inappro- cates’ down the newly synthesized G-rich strand, to
priate fusion. Although a human telomere binding pro- reanneal at the terminus where another telomere repeat
tein has been identified,2 the DNA-protein structure at is synthesized and so the process continues.I0J’ In this
the terminus of human telomeres is unknown. Telomeres manner, telomeres and telomerase are able to compen-
have also evolved to tolerate the sequence loss that arises sate for the sequence loss that arises from incomplete
from incomplete replication during S-phase of mitosis or replication.
meiosis. In humans, telomeres are composed of head to Telomere attrition in normal (primary) cultured
tail arrays of a short repeat unit, TTAGGG, though human cells correlates with the eventual senescence
many variant repeats occur at the proximal end of the of the culture after a finite number of population
array^.^ The length of the repeat array varies consider- doublings. The senescent cells show an increased
ably between telomeres, both within and between cells. frequency of chromosome breaks, fragments, and dicen-
Telomere repeat arrays are devoid of restriction endo- trics that arise from end to end fusions, all characteristic
nuclease cleavage sites, so digestion of genomic DNA of chromosomes that lack functional telomeres. It has
with almost any restriction enzyme releases the terminal been proposed that telomere length can be used as an
restriction fragment (TRF), which will include the tel- hdicator of replicative capacity in human somatic cells.
omere from each chromosome, by cleaving in the proxi- In agreement with this hypothesis, telomere length is
mal flanking DNA. The TRFs can then be detected shorter in the majority of colonic carcinomas than in the
as smears of TTAGGG hybridizing fragments between surrounding normal tissue, as a result of the mitotic
2 and 20 k ~ b . ~ clonal expansion that gave rise to the carcinoma.
The average telomere length varies between individ- Human telomerase activity was first detected in an
uals, but is similar between twins. The high heritability immortalized somatic cervical carcinoma cell line’ that
of telomere length (78 per cent) suggests that genetic has a short but stable telomere length. Recently a very
factors play a major role in its contr01.~In somatic sensitive assay for telomerase activity has shown that
tissues, the average telomere length decreased by about 98/100 immortalized cell lines and 90/101 samples from
30-50 bp per year, reflecting the gradual loss of 12 different tuhour types had telomerase activity,
sequences as a result of incomplete replication. The whereas no telomerase activity was detected in 50 nor-
gradual attrition of telomere length in somatic tissues mal tissue samples or 22 normal cell lines.12
correlates with increasing age and has been directly All of these observations lend support to what has
related to the number of cell divisions in cultures of become known as the telomere hypothesis of cellular
normal cell^.^.^ In trisomy 21 patients, telomere length senescence and imm~rtalization.’~ This proposes that
decreases more rapidly than in age-matched donors, telomerase is active in the germline, to maintain tel-
and in a premature ageing disorder, Hutchinson- omere length and replicative capacity in the gametes and
Gilford progeria, telomeres are also shorter, though hence for the next generation, but at some stage early in
the molecular basis for the increased rate of telomere embryogenesis telomerase activity is switched off or
attrition is not understood.* In the germline, however, repressed in the differentiating somatic tissues. As a
the average telomere length is stable and may increase result, telomere length decreases at each cell division and
slightly with increasing age, the sequence loss that though the average rate of decline can be measured, the
occurs during replication being compensated for by the actual sequence loss at each telomere may vary dramati-
activity of an unusual ribonucleoprotein called cally from cell to cell. After a finite number of divisions,
telomerase. the cells enter senescence. The counting mechanism for
Telomerase activity was first detected in the lower the number of cell divisions and the signal for entry into
eukaryote Tetrahymena, as an enzyme that synthesized senescence are unknown, but may involve telomere
the de novo addition of a telomere repeat array onto the length, such that short telomeres signal a checkpoint
end of a synthetic oligonucleotide. The enzyme has at arrest. However, if a cell bypasses the checkpoint, which
least two protein components and an RNA molecule has been achieved in culture by the expression of certain
CCC 0022-3417/96/110233-03 Received 15 April 1996
0 1996 by John Wiley & Sons, Ltd. Accepted 1 May 1996
234 REVIEW

viral proteins, and continues to divide, telomeres con- capacity.22 The limited replicative capacity is a particu-
tinue to shorten until a crisis is reached (average tel- lar problem in tissues, such as the blood, that have a
omere length is l.S kbp14). At this point, many very high turnover of cells throughout the life time of the
chromosomes lack functional telomeres and the cell will individual. While the teloinere length in blood DNA
die unless the telomere length is stabilized by the reacti- clearly decreases with increasing age, recent improve-
vation of telomerase. ments in the detection of telomerase activity indicate
The autosomal recessive disorder ataxia telangiectasia that some of the bone marrow stem cells express
is characterized by progressive cerebral ataxia, hyper- telomerase activity.23
sensitivity to ionizing radiation and an increased predis- A proportion of colonic carcinomas and tumours of
position to leukaemias and lymphomas. About 1 per other tissues have been identified where the telomeres
cent of the population, with a 3 to 4-fold increased risk are considerably longer than the surrounding normal
for predisposition to cancer, are heterozygous carriers tissues, but telomerase activity has not been detected.
for AT gene mutations. Cell lines from AT patients Recent investigation of virus transformed immortal cell
show reduced replicative capacity in culture, high levels cultures has shown that very long telomeres (approxi-
of chromosome instability, and frequent end to end mately 50 kbp) can be generated and maintained in the
chromosome associations, all of which indicate defects absence of telomerase activity.24 The mechanism for this
in telomere maintenance. The ATM gene shows strong is as yet unknown. An alternative pathway has been
homology to the functionally related yeast Tell and identified for yeast telomere maintenance, involving
Mecl genes, that are involved in a checkpoint arrest exchanges between sub-terminal repeats, and a similar
in response to DNA damage.lS.l6 Interestingly, tell process may rescue telomere length in some human
mutant yeast has shortened telomeres. AT cells are tumours. Studies of the distribution and variation of
probably defective in their ability to delay or arrest adjacent or sub-terminal repeat sequences of human
the cell cycle following DNA damage and given the telomeres indicate that non-homologous exchanges
apparent telomere defects observed in these cells, ATM in the distal region of chromosomes occur at a low
may play a role in the checkpoint initiated by shortened rate.2s
telomeres. l 7 The contribution of telomere loss and telomerase
If, as seems likely, telomeres and telomerase play a activity has been considered for diseases of somatic
role in signalling cellular senescence and in the progres- tissues but there is evidence that chromosome breaks in
sion of tumorigenesis, anti-cancer therapies could be the germline are sometimes healed inappropriately by
targeted at telomerase. The gene encoding the human telomerase. Telomerase has relatively few sequence
RNA component of telomerase has recently been requirements before it is able to seed the formation of a
cloned; the RNA transcript, hTR, is about 450 bases new telomere onto the end of any DNA molecule, so
with a template region of 11 nucleotides (S’- ‘capping’ the chromosome and preventing further
CUAACCCUAAC). I x Encouragingly, expression of genome imbalance. The deleted chromosome may arise
anti-sense to hTR in an immortal telomerase-expressing in a gamete, but as the majority of terminal deletions are
cell line resulted in a gradual reduction of telornere incompatible with life, only the smaller and so-called
length, leading to death of the cells after about 26 ‘cryptic’ deletions tend to occur in patients. These
population doublings.l8 Other studies, though, using patients present with features of a contiguous gene
reverse transcriptase inhibitors in immortal T- and B-cell deletion syndrome, usually including mental retarda-
lines, showed that the telomere length was highly unsta- tion. A few terminal deletion breakpoints from patients
ble and that another, telomerase-independent, mechan- with a-thalassaemia have been cloned, each capped by
ism could contribute to telomere maintenance.I9 As the presence of a novel telomere.26
expected, hTR has been detected in germline and in In summary, the loss of a telomere through the
tumour tissues that express telomerase activity; unex- gradual attrition of the repeat array (or DNA damage)
pectedly, the RNA component was also detected, but at leads to extreme genome instability. Consequently it
a lower level, in sonie somatic tissues that do not express probably triggers a somatic cell cycle arrest, perhaps
the enzyme activity,18 indicating that the regulation of mediated in part by the ATM protein, and entry into
the protein and RNA components of human telomerase senescence. The maintenance of functional telomeres is
are different. required, but is probably not sufficient for somatic cell
Recent studies of the timing of reactivation of telom- immortality. Defects in any of the processes that main-
erase in pre-neoplastic and neoplastic gastric and colo- tain telomere integrity, control entry into senescence, or
rectal lesions and in prostate tissues indicate that it regulate telomerase repression are therefore likely to
usually occurs early in carcinogenesis. However, the contribute to disease. In the germline, telomerase main-
variation in telomere lengths between tumours suggests tains telomere length and chromosome integrity for the
that the timing of reactivation is not precise.20.21The next generation, but inappropriate healing of chromo-
telomerase-expressing malignant cells are able to main- some breaks also contributes to gametic imbalance and
tain telomere length arid genome composition and may disease.
therefore have a growth advantage over other non-
expressing malignant cells and outgrow them. It has NICOLA J. ROYLE
been proposed that the control of telomerase activity in Department of Genetic.,
humans gives added protection against the multistep University of Leicester
process of tumorigenesis by limiting replicative Leicester LEI 7RH, U.K.
 REVIEW 235

REFERENCES 15. Morrow DM, Tagle DA, Shiloh Y, Collins FC, Hieter P. TELI, an S.
cerevisiue homolog of the human gene mutated in ataxia telangiectasia, is
1. Zakian VA. Telomeres: beginning to understand the end. SciencuA995; 270 functionally related to the yeast checkpoint gene MECI. Cell 1995; 8 2
1601-1 606. s3 1-840.
2. Chong L, van Steensel B, Broccoli D, er a / . A human telomeric protein. 16. Greenwell PW, Kronmal SL, Porter SE, e f ul. TELI, a gene involved in
Science 1995; 270 1663-1667. controlling telomere length in S. cerevisiae, is homologous to the human
3. Baird DM, Jeffreys AJ, Roy1 NJ. Mechanisms underlying telomere repeat ataxia telangiectasia gene. Cell 1995; 82: 823-829.
turnover, revealed by hypervariable variant repeat distribution patterns in 17. Sanchez Y, Desany BA, Jones WJ, e f ul. Regulation of RAD53 by the
the huinan Xp/Yp telomere. E M 5 0 J 1995; 1 4 ?. ATM-like kinases MECI and TELI in yeast cell cycle checkpoint pathwaya.
4. Moyzis RK, Buckingham JM, Cram LS, ef ul. A highly repetitive DNA Science 1996; 271: 357-360.
sequence, (TTAGGG),,, present at the telomeres of human chromosomes. 18. Feng J, Funk WD, Wang S-S, el ul. The RNA component of human
Pro< Acud Sci USA 1988; 8 5 6622-6626. telomerase. Science 1995; 269 12361241.
5. Slagbloom PE, Droog S, Boomsma DI. Genetic determination of telomere 19. Strahl C, Blackburn EH. Effects of reverse-transcriptase inhibitors on
size in humans: a twin study of three age groups. A m J Hum Genet 1994; 55: telomere length and telomerase activity in two immortalized human cell-
876-882. lines. Mol Ce/l B i d 1996; 1 6 53-65.
6. Hastie ND, Dempster M, Dunlop MG, ef a/. Telomere reduction in human 20. Tahara H, Kuniyasu H, Yokozaki H, ef ul. Telomerase activity in preneo-
colorectal carcinoma and with ageing. Nuture 1990, 346 866-868. plastic and neoplastic gastric and colorectal lesio Clin Cuncer Rev 1995; 1:
7. Harley CB, Futcher AB, Greider CW. Telomeres shorten during ageing of
human fibroblasts. Nafurc 1990; 345 458460. 21.
1245-125 I 7-
Sommerfeld HJ, Meeker AK, Piatyszek MA, e f ul. Telomerase activity-~-a
8. Vaziri H, Schachter F, Uchida I, ef ul. Loss of telomeric DNA during aging prevalent marker of malignant human prostate tissue. Cancer Rrs 1996; 5 6
of normal and trisomy 21 human lymphocytes. Am J Hum Genet 1993; 52: 2 18-222.
661-667. 22. Prowse KR, Greider CW. Developmental and tissue-specific regulation of
9. Collins K, Kobayashi R, Greider CW. Purification of Tet?’uhji‘mrnatelom- mouse telomerase and telomere length. Proc Nurl Acud Sri 1995; 9 2
erase and cloning of genes encoding the two protein compoiients of the 48 184822.
enzyme. Cell 1995; 81: 677-686.
23. Broccoli D, Young JW, de Lange T. Telomerase activity in normal and
10. Greider CW, Blackburn EH. A telomeric sequence in the RNA of TcJtruhy-
IZEIUZtelomerase required for telomere repeat synthesis. Nuture 1989; 337:
malignant hematopoietic cells. Proc Natl Acud Sci USA 1995; 9 2 9082-
331-337. 9086.
11. Morin GB. The human telomere terminal transferase enzyme is a ribo- 24. Bryan TM, Englezou A, Gupta J , Bacchetti S , Reddel RR. Telomere
nucleoprotein that synthesizes TTAGGG repeats. Cell 1989; 5 9 521-529. elongation in immortal human cells without detectable telomerase activity.
12. Kim NW, Piatyszek MA, Prowse KR, r t ul. Specific association of human EMBO J 1995; 1 4 42404248.
’
telomerase activity with immortal cells and cancer. Science 1994; 2 6 6 25. Wilkie AO, Higgs DR, Rack KA, ef ul. Stable length polymorphism of up
2011-2015. to 260 kb at the tip of the short arm of human chromosome 16. Cell 1991;
13. Greider CW. Mammalian telomere dynamics: healing, fragmentation short- 64: 595-606.
ening and stabilization. Curr Op Genef Dev 1994; 4 203-211. 26. Flint J, Craddock CF, Villegas A, et al. Healing of broken human
14. Counter CM, Avilion AA, LeFeuvre CE, ef ul. Telomere shortening chromosomes by the addition of telomeric repeats. Am J Hum Genet 1994;
associated with chromosome insrability is arrested in immortal cells which 55: 505-512.
express telomerase activity. E M B O J 1992: 11: 1921-1929.