Upstream eventIncrease, Chromosomal aberrations
Increase, Cell Proliferation
Key Event Relationship Overview
AOPs Referencing Relationship
|AOP Name||Adjacency||Weight of Evidence||Quantitative Understanding|
|Direct deposition of ionizing energy leading to lung cancer||adjacent||Moderate||Low|
Life Stage Applicability
|All life stages||High|
Key Event Relationship Description
CAs are defined as abnormalities in the chromosome structure, often due to losses or gains of chromosome sections or the entire chromosomes itself, or chromosomal rearrangements (van Gent et al., 2001). These aberrant structures can come in a multitude of different forms. Types of CAs include: inversions, insertions, deletions, translocations, dicentric chromosomes (chromosomes that contain two centromeres, often resulting from telomere end fusions (Fenech & Natarajan 2011; Rode et al., 2016), centric ring chromosomes, acentric chromosome fragments, micronuclei (MN; small nucleus-like structures containing entire chromosomes or chromosome fragments (Fenech & Natarajan, 2011; Doherty et al., 2016), nucleoplasmic bridges (NBPs; a corridor of nucleoplasmic material containing chromatin that is attached to both daughter cell nuclei), nuclear buds (NBUDs; small MN-type structures that are still connected to the main nucleus (Fenech & Natarajan, 2011), and copy number variants (CNVs; deletions or duplications of chromosome segments (Russo et al., 2015).
If these CAs affect genes involved in controlling the cell cycle, this may result in increased cellular proliferation. There are three types of genes that, if modified, may result in high rates of proliferation: proto-oncogenes, tumour suppressor genes (TSGs), and caretaker/stability genes (Vogelstein & Kinzler, 2004; Hanahan & Weinberg, 2011). Furthermore, gene fusions that result from CAs have also been implicated in augmenting cellular proliferation (Sanders & Albitar, 2010; Ghazavi et al., 2015; Kang et al., 2016).
Evidence Supporting this KER
There is a strong biological plausibility for a relationship between CAs and rates of cellular proliferation. This is particularly emphasized in the context of carcinogenesis, as high cellular proliferation is a known hallmark of cancer, and an enabling characteristic of increased proliferation is genomic instability (Hanahan & Weinberg, 2011).Topical reviews are available documenting the contribution of CAs to cellular proliferation and/or cancer development (Mes-Masson & Witte, 1987; Bertram, 2001; Vogelstein & Kinzler, 2004; Ghazavi et al. ,2015; Kang et al., 2016). The link between chromosomal instability (CIN), which describes the rate of chromosome gains and losses, and cancer development has also been well documented (Thompson et al., 2017; Gronroos, 2018; Targa & Rancati, 2018; Lepage et al., 2019).
Many CAs are thought to be formed through two main mechanisms: inadequate repair of DNA damage, and errors in mitosis. If there is damage to the DNA that the cell is unable to properly repair, the unrepaired lesion may translate into a CAs (Bignold, 2009; Danford, 2012; Schipler & Iliakis, 2013); the type of resulting CA is often influenced by the cell cycle stage when the damage occurred (Danford, 2012; Registre et al., 2016; Vodicka et al., 2018), and the type of erroneous repair (Ferguson & Alt, 2001; Povirk, 2006; Bignold, 2009; Danford, 2012; Schipler & Iliakis, 2013). Errors made during repair may be particularly detrimental if they interrupt or modify critical genes, or if chromosome structures are created that cannot undergo mitosis (Schipler & Iliakis, 2013). Similarly, errors in mitosis that prevent chromosomes from being properly segregated may also lead to CAs. These errors could be due to by improper timing of centrosome separation, the presence of extra centrosomes, inappropriate mitotic spindle assembly and attachment to kinetochores (found on the centromeres), and incorrect sister-chromatid cohesion (Levine & Holland, 2018).
The presence of CAs in cells may be particularly detrimental if they alter the rate of cellular proliferation by affecting genes that control the cell cycle, namely proto-oncogenes, TSGs (Bertram, 2001; Vogelstein & Kinzler, 2004) or caretaker/stability genes (Vogelstein & Kinzler, 2004). Proto-oncogenes are genes that, when activated, promote cellular proliferation. CAs that increase activation of these genes may aberrantly boost cell cycling and therefore increase proliferation (Bertram, 2001; Vogelstein & Kinzler, 2004). Activation of proto-oncogenes have also been implicated in the cancer stem cell theory of carcinogenesis (Vicente-duen et al., 2013). Examples or proto-oncogenes include EGFR and KRAS (Sanders & Albitar, 2010). TSGs refer to genes that actively suppress cell proliferation and, in some cases, promote apoptosis (Bertram, 2001; Vogelstein & Kinzler, 2004; Sanders & Albitar, 2010). If these genes are silenced by CAs, this may remove cell cycle checkpoints, thus allowing for unhindered cellular proliferation and decreased apoptosis (Bertram, 2001; Vogelstein & Kinzler, 2004). Common TSGs are TP53 and RB (Hanahan & Weinberg, 2011). Lastly, caretaker/stability genes are those involved in the prevention and detection of DNA damage, and the instigation and completion of the required DNA repair (Vogelstein & Kinzler, 2004; Hanahan & Weinberg, 2011). If the function of these caretaker/stability genes is affected by CAs, this may result in genome-wide inadequate DNA repair, which in turn may result in genetic damage to TSGs or proto-oncogenes (Vogelstein & Kinzler, 2004). Genes involved in mismatch repair (MMR), nucleotide-excision repair (NER) and base-excision repair (BER) are all examples of caretaker/stability genes (Vogelstein & Kinzler, 2004).
There are also other CAs commonly associated with cancer. In prostate cancer, truncated TSGs such as TP53, PTEN, BRCA1, and BRCA2 are a result of chromosomal rearrangements (Mao et al., 2011). Similarly, chromosomal inversions were found to be responsible for just over half of the RET gene fusions associated with lung adenocarcinoma samples (Mizukami et al., 2014).
There is moderate empirical evidence supporting the relationship between CAs and the cellular proliferation. The evidence presented below is summarized in table 8, here (click link). There are some available reviews that provide evidence for this relationship in the context of carcinogenesis, as high levels of cellular proliferation is one of the hallmarks of cancer (Hanahan & Weinberg, 2011). Many of these reviews focus especially on the structure and function of specific cancer-associated CAs (Mes-Masson & Witte, 1987; Ghazavi et al., 2015; Kang et al., 2016). Another interesting review discusses transgenic mouse models that have contributed to our understanding of how oncogenes and TSGs promote carcinogenesis in a variety of tissues (Fowlis & Balmain, 1992). Overall, however, there is a lack of empirical evidence available supporting dose and incidence concordance, little empirical evidence supporting temporal concordance, but strong empirical evidence supporting essentiality for this KER.
Dose and Incidence Concordance
There were no studies identified that directly assessed the temporal concordance between CA and increasing rates of cellular proliferation. In a study examining MN frequency and cell proliferation in estrogen-responsive cancer cells treated with estradiol, both MN levels and proliferation rates were higher in estradiol-treated cells relative to controls at 140 and 216 hours post-treatment (Stopper et al., 2003). This suggests that both events are increased at the same time points in response to the estradiol. More work is required, however, to directly assess the temporal concordance between CA frequency and cell proliferation rates.
Much of the evidence for essentiality stems from studies of gene fusions produced by chromosomal translocations and the corresponding impact on cellular proliferation rates. One such gene fusion, JAFZ1-JJAZ1, has been identified in endometrial stromal sarcomas. The role of this relatively unknown translocation was evaluated using knock-down and knock-in experiments. When wild-type JJAZ1 was disabled by siRNA, HEK 293 cells expressing the JAFZ1-JJAZ1 fusion were found to have an increased rate of cellular proliferation (Li et al., 2007). Similarly, the role of the EML4-ALK fusion gene was examined in IL-3 dependent BA/F3 cells. These cells were transfected with a plasmid carrying only CD8, or CD8 in combination with ALK, EML4-ALK, or mutant EML4-ALK (which contained a lysine to methionine mutation in the kinase domain). In all cases, cell proliferation was found to increase linearly over 7 days in the presence of IL-3; in the absence of IL-3, all cells died by day 3 of culture, with the exception, however, of cells carrying EML4-ALK. Only cells with EML4-ALK were able to maintain a positive, linear growth in both the presence and absence of IL-3. Addition of a JAK2 inhibitor to these EML4-ALK cells resulted in a dose-dependent decline in cellular proliferation, such that at a dose of 10 um of inhibitor, cells numbers declined steadily until death at day 5. This is in contrast to the CD8-expressing cells exposed to the same inhibitor doses, in which there was only a very slight decline in cellular proliferation rates (Soda et al. 2007). Both of these studies provide evidence that translocations increase proliferation rates in cells.
In addition to causing gene fusions, translocations may also lead to the production of circular RNA fusion products (f-CircRNA), which can be studied to further understand the link between CAs and cellular proliferation. For example, f-CircPR has been associated with the PML-RARα translocation, f-CircM9 has been associated with the MLL-AF9 translocation, and expressions of f-CircPR or f-CircM9 were both found to increase cell proliferation rates in mouse embryonic fibroblasts. Inhibition of these f-CircRNAs, either through addition of silencing shRNA or by using a mutant non-circularizing f-CircRNA, resulted in decreased rates of cell proliferation (Guarnerio et al., 2016). These results again indicate that there is a relationship between CAs and increased cellular proliferation.
Other experiments provide evidence that CAs can increase cellular proliferation using cancer cells. Using two human Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL) cell lines (both of which contain the BCR-ABL translocation), cellular proliferation was studied by cell counting and by analyzing levels of phosphorylated ErbB2. ErbB2 is a member of the ERB receptor tyrosine kinase family that is involved in pro-proliferative signalling, and it is known to be expressed in cells from ALL patients. Cell proliferation rates were found to decline in a dose-dependent fashion when treated with either an ErbB family tyrosine kinase inhibitor, or a more specific ErbB1/ErbB2 tyrosine kinase inhibitor. Furthermore, treatment with the ErbB family inhibitor also resulted in significant decreases in phosphorylated ErbB2 (Irwin et al., 2013). In another set of experiments using estrogen receptor-positive human ovarian cancer cells, treatment of cells with estrogen were found to have significantly increased levels of MN and significantly increased proliferation rates relative to vehicle-treated control cells; furthermore, there were more cells in S-phase and fewer in the G2/M phases of the cell cycle relative to controls. These results were specific to estrogen-response cells, as treatment of estrogen receptor-negative human ovarian cancer cells with estrogen did not result in any changes to MN or cell proliferation. Furthermore, addition of an estrogen antagonist to estrogen-responsive cells maintained MN frequencies and cell proliferation at control levels (Stopper et al., 2003).
Human Epidemiology Association and Genetic Studies
Very often, CAs result in gene fusions. A gene fusion occurs when two genes not normally in close proximity to each other are juxtaposed; this may result in altered expression of one or both genes, or an altered gene product (Mitelman, 2005). There are several well-known gene fusions implicated in carcinogenesis that are associated with increased cellular proliferation. One well-characterized gene fusion is the Philadelphia chromosome, also known as the BCR-ABL1 fusion. This gene fusion is formed by a translocation between chromosome 9 and 22, and is commonly found in chronic myelogenous leukemia (CML) (Mes-Masson and Witte 1987; Kang et al. 2016), as well as acute lymphoblastic leukemia (ALL) (Ghazavi et al., 2015). The protein created from BCR-ABL1 has elevated tyrosine kinase activity, and was shown to increase activation of cellular proliferation pathways (Ghazavi et al., 2015; Kang et al., 2016) including JAK2/STAT, PI3K-AKT, and MAPK/ERK (Kang et al., 2016). Another common gene fusion partner is ALK, which is a receptor tyrosine kinase involved in the PI3K-AKT signalling pathway. Very often, ALK gene fusions result in upregulated ALK expression, and a resulting increase in pro-proliferative signalling in the PI3K-AKT pathway. In non-small cell lung cancer, the ALK-EML4 gene fusion is particularly common (Sanders & Albitar, 2010). Similarly, ETV6-RUNX1 is the most common fusion gene in B-cell precursor acute lymphoblastic leukemia (BCP-ALL), and is thought to initiate leukemogenesis (Ghazavi et al., 2015).
Uncertainties and Inconsistencies
Uncertainties in this KER are as follows:
- A study using peripheral blood lymphocytes isolated from head and neck cancer patients found significantly increased CAs (including chromosome-type aberrations, chromatid-type aberrations, dicentric chromosomes, aneuploidy, MN, NPBs and NBUDs) relative to healthy controls. In the lymphocytes from these same cancer patients, however, the cell proliferation rates were significantly decreased (George et al., 2014).
- Characterization of 20 different ameloblastomas, which are benign tumours associated with the jaw, found low CAs frequencies and low rates of cellular proliferation (Jääskeläinen et al., 2002).
Quantitative Understanding of the Linkage
Quantitative understanding has not been well-established for this KER. There were no studies identified that documented a response-response relationship between CA frequency and cell proliferation rates, and a severe lack of time scale-oriented studies. Overall, more research is required to establish a quantitative understanding of this KER.
Studies that directly assessed the time scale between CAs and cellular proliferation were not identified. However, differences in cellular proliferation rates for cells with different CA-related manipulations or treatments were evident within the first 3 days of culture (Stopper et al., 2003; Li et al., 2007; Soda et al., 2007; Irwin et al., 2013; Guarnerio et al., 2016). More studies are required, however, to formulate a detailed time scale relating these two events.
Known modulating factors
Known Feedforward/Feedback loops influencing this KER
Domain of Applicability
The domain of applicability pertains to all multicellular organisms, as cell proliferation and death regulate tissue homeostasis (Pucci et al., 2000).
Bertram, J.S. (2001), "The molecular biology of cancer.", Mol. Aspects. Med. 21:166–223. doi:10.1016/S0098-2997(00)00007-8.
Bignold, L.P. (2009), "Mechanisms of clastogen-induced chromosomal aberrations: A critical review and description of a model based on failures of tethering of DNA strand ends to strand-breaking enzymes.", Mutat. Res., 681(2-3):271–298. doi:10.1016/j.mrrev.2008.11.004.
Danford, N. (2012), "The Interpretation and Analysis of Cytogenetic Data.", Methods Mol. Biol., 817:93-120. doi:10.1007/978-1-61779-421-6.
Doherty, A., S.M. Bryce & J.C. Bemis (2016), "The In Vitro Micronucleus Assay. Methods in molecular biology", (Clifton, N.J.). 817:121-41. doi: 10.1007/978-1-61779-421-6_7.
Fenech, M. & A.T. Natarajan (2011), "Molecular mechanisms of micronucleus, nucleoplasmic bridge and nuclear bud formation in mammalian and human cells.", Mutagenesis 26(1):125–132. doi:10.1093/mutage/geq052.
Ferguson, D.O. & F.W. Alt (2001), "DNA double strand break repair and chromosomal translocation: Lessons from animal models.", Oncogene 20(40):5572–5579.
Fowlis, D.J. & A. Balmain (1993), "Oncogenes and Tumour Suppressor Genes in Transgenic Mouse Models of Neoplasia.", Eur. J. of Cancer 29A(4):638-45. doi: 10.1016/S0959-8049(05)80170-4.
van Gent D.C., J.H.J. Hoeijmakers & R. Kanaar (2001), "Chromosomal stability and the DNA double-stranded break connection.", Nat. Rev. Genet. 2(3):196–206. doi:10.1038/35056049. http://www.ncbi.nlm.nih.gov/pubmed/11256071.
George, A., R. Dey & V.B. Dqhumhh (2014), "Nuclear Anomalies, Chromosomal Aberrations and Proliferation Rates in Cultured Lymphocytes of Head and Neck Cancer Patients.", Asian Pacific journal of cancer prevention. 15(3):1119-1123. doi:10.7314/APJCP.2014.15.3.1119.
Ghazavi, F. et al. (2015), "Molecular basis and clinical significance of genetic aberrations in B-cell precursor acute lymphoblastic leukemia.", Exp Hematol. 43(8):640–653. doi:10.1016/j.exphem.2015.05.015.
Gronroos, E. (2018), "Tolerance of Chromosomal Instability in Cancer: Mechanisms and Therapeutic Opportunities.", Cancer Res. 78(23):6529-6535, doi:10.1158/0008-5472.CAN-18-1958.
Guarnerio, J. et al. (2016), "Oncogenic Role of Fusion-circRNAs Derived from Article Oncogenic Role of Fusion-circRNAs Derived from Cancer-Associated Chromosomal Translocations.", Cell. 165(2):289–302. doi:10.1016/j.cell.2016.03.020.
Hanahan, D. & R.A. Weinberg (2011), "Review Hallmarks of Cancer: The Next Generation.", Cell. 144(5):646–674. doi:10.1016/j.cell.2011.02.013.
Irwin, M.E. et al. (2013), "Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome – Positive Acute Lymphoblastic Leukemia.", PLoS One, 8(8):1–10. doi:10.1371/journal.pone.0070608.
Jääskeläinen, K. et al. (2002), "Cell proliferation and chromosomal changes in human ameloblastoma.", Cancer Genetics and Cytogenetics. 136(1):31-7. doi: 10.1016/S0165-4608(02)00512-5.
Kang, Z.J. et al. (2016), "The Philadelphia chromosome in leukemogenesis.", Chin J Cancer.:1–15. doi:10.1186/s40880-016-0108-0.
Lepage, C.C. et al. (2019), "Detecting Chromosome Instability in Cancer: Approaches to Resolve Cell-to-Cell Heterogeneity.", Cancers (Basel), 11(2): pii: E226. doi:10.3390/cancers11020226.
Levine, M.S. & A.J. Holland (2018), "The impact of mitotic errors on cell proliferation and tumorigenesis.", Genes Dev., 32(9-10):620–638. doi:10.1101/gad.314351.118.620.
Li, H. et al. (2007), "Effects of rearrangement and allelic exclusion of JJAZ1 / SUZ12 on cell proliferation and survival.", PNAS, 104(50):20001–20006.
Mao, X. et al. (2011), "Chromosome rearrangement associated inactivation of tumour suppressor genes in prostate cancer.", American Journal of Cancer Research. 1(5):604-17.
Mes-Masson, A.-M. & O.N. Witte (1987), "Role of The abl Oncogene in Chronic Myelogenous Leukemia.", Advances in Cancer Research. 49:53-74. doi: 10.1016/S0065-230X(08)60794-0.
Mitelman, F. (2005), "Deep Insight Section: Cancer cytogenetics update", Atlas of Genetic and Cytogenetics in Oncology and Haematology, 9(2):188–190. doi:10.4267/2042/38202.
Mizukami, T. et al. (2014), "Molecular Mechanisms Underlying Oncogenic RET Fusion in lung adenocarcinoma.", J Thorac Oncol. 9(5):622–630. doi:10.1097/JTO.0000000000000135.
Povirk, L.F. (2006), "Biochemical mechanisms of chromosomal translocations resulting from DNA double-strand breaks.", DNA Repair (Amst). 5(9-10):1199–1212. doi:10.1016/j.dnarep.2006.05.016.
Pucci, B., M. Kasten & A. Giordano (2000), "Cell Cycle and Apoptosis 1", Neoplasia, 2(4):291–299.doi: 10.1038/sj.neo.7900101
Registre, M., R. Proudlock & N. Carolina (2016), "The In Vitro Chromosome Aberration Test.", Genetic Toxicology Testing. 207-267. doi: 10.1016/B978-0-12-800764-8.00007-0.
Rode, A. et al. (2016), "Chromothripsis in cancer cells: An update.", Int. J. Cancer, 138(10):2322–2333. doi:10.1002/ijc.29888.
Russo, A. et al. (2015), "Review Article Genomic Instability: Crossing Pathways at the Origin of Structural and Numerical Chromosome Changes.", Envrion. Mol. Mutagen. 56(7):563-580. doi:10.1002/em.
Sanders, H.R. & M. Albitar (2010), "Somatic mutations of signaling genes in non-small-cell lung cancer.", Cancer Genet Cytogenet. 203(1):7–15. doi:10.1016/j.cancergencyto.2010.07.134.
Schipler, A. & G. Iliakis (2013), "DNA double-strand – break complexity levels and their possible contributions to the probability for error-prone processing and repair pathway choice.", Nucleic Acids Res., 41(16):7589–7605. doi:10.1093/nar/gkt556.
Soda, M. et al. (2007), "Identification of the transforming EML4 – ALK fusion gene in non-small-cell lung cancer.", Nature, 448(7153):561-566. doi:10.1038/nature05945.
Stopper, H. et al. (2003), "Increased cell proliferation is associated with genomic instability: elevated micronuclei frequencies in estradiol-treated human ovarian cancer cells.", Mutagenesis 18(3):243-247. doi:10.1093/mutage/18.3.243.
Targa, A. & G. Rancati (2018), "Cancer: a CINful evolution.", Curr Opin Cell Biol. 2018 Jun;52:136-144., doi:10.1016/j.ceb.2018.03.007
Thompson, L.L. et al. (2017), "Evolving Therapeutic Strategies to Exploit Chromosome Instability in Cancer.", Cancers (Basel), 9(11): pii: E151 doi:10.3390/cancers9110151.
Vicente-Duenas, C. et al. (2013), "Function of oncogenes in cancer development: a changing paradigm", EMBO J., 32(11):1502–1513. doi:10.1038/emboj.2013.97.
Vodicka, P. et al. (2018), "Genetic variation of acquired structural chromosomal aberrations.", Mutat Res Gen Tox En, 836(May):13–21. doi:10.1016/j.mrgentox.2018.05.014.
Vogelstein, B. & K.W. Kinzler (2004), "Cancer genes and the pathways they control.", Nat. Med, 10(8):789–799. doi:10.1038/nm1087.