TPX2 Amplification-Driven Aberrant Mitosis in Culture Adapted Human Embryonic Stem Cells with gain of 20q11.21

Jeong, Ho-Chang; Go, Young-Hyun; Shin, Joong‐Gon; Kim, Yun-Jeong; Cho, Min-Guk; Gwon, Dasom; Cheong, Hyun Sub; Lee, Haeseung; Lee, Jae-Ho; Jang, Chang-Young; Shin, Hyoung Doo; Cha, Hyuk‐Jin

doi:10.1007/s12015-023-10514-4

Cited by 4 publications

(9 citation statements)

References 49 publications

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“…These results suggest that an epigenetic change that alters chromatin structures regulating genes associated with 'spindle organization' is induced in culture-adapted variants (i.e., P3 and P4). Consistently, the aberrant mitosis with a lagging chromosome or chromosome bridge that results from abnormal spindle dynamics occurs in these cultureadapted variants (Jeong et al, 2023).…”

Section: Alteration Of Chromatin Accessibility In Culture-adapted Var...mentioning

confidence: 66%

“…Genomic variants (i.e., P3 and P4 hESCs) share typical culture-adapted phenotypes, as previously demonstrated (Avery et al ., 2013; Markouli et al , 2019; Nguyen et al ., 2014; Price et al , 2021; Spits et al , 2008; Zhang et al ., 2019) and depicted in Figure 1A. As these culture-adapted phenotypes were manifested in P3 hESCs (but not P2 hESCs), P4 hESCs with an additional 17q24 gain (Jeong et al ., 2023) (Fig. S1A) from P3 hESCs (with a 20q11.21 gain) encourages closer examination of ‘stepwise variation models for genome hESCs instability (Ben-David, 2015; Halliwell et al ., 2020).…”

Section: Resultsmentioning

confidence: 99%

“…We hypothesized that a molecular event to activate the chromatin structure of 20q11.21 loci would transpire in hPSCs along with a 20q11.21 gain. This event would also promote crucial gene expressions, such as BCL2L1 (for survival) or TPX2 (for spindle stabilization and abnormal mitosis), to achieve the typical cellular phenotypes of culture-adapted variants (i.e., survival trait and abnormal mitosis (Avery et al ., 2013; Jeong et al ., 2023; Nguyen et al ., 2014) (Fig. 5A).…”

Section: Resultsmentioning

confidence: 99%

“…The recent meta-analysis of 107 different studies revealed that chromosomes 20, 12, 17, X, and 1 are prevalent hotspots for genetic abnormalities (Assou et al , 2020). Despite emerging reports on altered cellular phenotypes, such as acquired survival traits (Avery et al ., 2013; Cho et al ., 2018; Nguyen et al ., 2014), aberrant mitosis (Jeong et al ., 2023), aneuploidy (Ben-David et al ., 2014; Na et al , 2014), and tumor development (Moon et al , 2011; Yamamoto et al , 2022), the underlying molecular processes for acquiring “culture-adapted phenotypes” or minimum risk assessments remain poorly understood.…”

Section: Discussionmentioning

confidence: 99%

“…This observation may explain the frequent 20q11.21 loci gains containing BCL2L1 (International Stem Cell et al , 2011) to induce anti-apoptotic BCL-xL protein (Avery et al , 2013; Cho et al , 2018; Nguyen et al , 2014) and 17p13.1 loss where TP53 is present (Amir et al , 2017) and somatic TP53 mutations (Merkle et al , 2017) in hPSCs during prolonged in vitro culture. Interestingly, these genetic aberrations such as the induction of BCL-xL as an ‘acquired survival trait’, serve to rescue cells from apoptosis triggered by aberrant mitosis, often resulting from TPX2 induction in hPSCs (Jeong et al , 2023). This, in turn, lead to aneuploidy or additional aberrations (Zhang et al , 2019).…”

Section: Introductionmentioning

confidence: 99%

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Sequential genetic and epigenetic alterations in human pluripotent stem cells for recurrent abnormality

Kim,

Kang,

Kweon

et al. 2023

Preprint

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Human embryonic stem cells (hESCs) are naturally equipped to maintain genome integrity to minimize genetic mutations during early embryo development. However, genetic aberration risks and subsequent cellular changes in hESCs duringin vitroculture pose a significant threat to stem cell therapy. While a few studies have reported specific somatic mutations and copy number variations (CNVs), the molecular mechanisms underlying ‘culture-adapted phenotype’ acquisitions of hESCs are largely unknown. Therefore, we conducted comprehensive genomic, single-cell transcriptomic, and single-cell ATAC-seq analyses of an isogenic hESC model displaying definitive ‘culture-adapted phenotypes.’ Notably, hPSCs with a copy number gain of 20q11.21 during early passage did not present ‘culture-adapted phenotypes’ norBCL2L1induction. Single-cell RNAseq and ATACseq analyses revealed active transcriptional regulation at 20q11.21 loci at late-passaged hESCs with the inducedBCL2L1andTPX2to trigger ‘culture-adapted phenotypes’ was associated with epigenetic changes facilitating TEA domain (TEAD) binding. These results suggest that copy number 20q11.21 gain and additional epigenetic changes are necessary for expressing ‘culture-adapted phenotypes’ by activating gene transcription at this specific locus.

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Section: Alteration Of Chromatin Accessibility In Culture-adapted Var...mentioning

confidence: 66%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Sequential genetic and epigenetic alterations in human pluripotent stem cells for recurrent abnormality

Kim,

Kang,

Kweon

et al. 2023

Preprint

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Safety Issues Related to Pluripotent Stem Cell-Based Therapies: Tumour Risk

Hillenius,

Montilla-Rojo,

Eleveld

et al. 2023

Pluripotent Stem Cell Therapy for Diabetes

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Epigenetic repression of CHCHD2 enhances survival from single cell dissociation through attenuated Rho A kinase activity

Kim,

Kwon,

Kim

et al. 2024

Cell. Mol. Life Sci.

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During in vitro culture, human pluripotent stem cells (hPSCs) often acquire survival advantages characterized by decreased susceptibility to mitochondrial cell death, known as “culture adaptation.” This adaptation is associated with genetic and epigenetic abnormalities, including TP53 mutations, copy number variations, trisomy, and methylation changes. Understanding the molecular mechanisms underlying this acquired survival advantage is crucial for safe hPSC-based cell therapies. Through transcriptome and methylome analysis, we discovered that the epigenetic repression of CHCHD2, a mitochondrial protein, is a common occurrence during in vitro culture using enzymatic dissociation. We confirmed this finding through genetic perturbation and reconstitution experiments in normal human embryonic stem cells (hESCs). Loss of CHCHD2 expression conferred resistance to single cell dissociation-induced cell death, a common stress encountered during in vitro culture. Importantly, we found that the downregulation of CHCHD2 significantly attenuates the activity of Rho-associated protein kinase (ROCK), which is responsible for inducing single cell death in hESCs. This suggests that hESCs may survive routine enzyme-based cell dissociation by downregulating CHCHD2 and thereby attenuating ROCK activity. These findings provide insights into the mechanisms by which hPSCs acquire survival advantages and adapt to in vitro culture conditions.

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TPX2 Amplification-Driven Aberrant Mitosis in Culture Adapted Human Embryonic Stem Cells with gain of 20q11.21

Cited by 4 publications

References 49 publications

Sequential genetic and epigenetic alterations in human pluripotent stem cells for recurrent abnormality

Sequential genetic and epigenetic alterations in human pluripotent stem cells for recurrent abnormality

Safety Issues Related to Pluripotent Stem Cell-Based Therapies: Tumour Risk

Epigenetic repression of CHCHD2 enhances survival from single cell dissociation through attenuated Rho A kinase activity

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