KR-20260062176-A - Biomarker composition for predicting stem cell-derived cardiomyocyte differentiation potential containing TEK and SDR42E1 genes and method for predicting stem cell-derived cardiomyocyte differentiation potential through measurement of TEK and SDR42E1

Abstract

The present invention relates to a biomarker composition for predicting the differentiation potential of stem cells into cardiomyocytes, comprising as an active ingredient a preparation for measuring the expression or activity level of TEK and SDR42E1 genes or proteins, and a method for predicting the differentiation potential of stem cells into cardiomyocytes through the measurement of TEK and SDR42E1. In the present invention, it was confirmed that the expression of TEK and SDR42E1 genes is evidenced in induced pluripotent stem cells that have a low degree of differentiation into cardiomyocytes, i.e., instability. Through this, it was confirmed that the stability of polyclonal cells can be predicted through the difference in expression of the two genes (TEK and SDR42E1). Accordingly, it is expected that the high cost/low efficiency problems that were issues in the commercialization of stem cells can be overcome through the preemptive stability evaluation technology of the present invention, and that the level of stem cell technology will be improved.

Inventors

• 이재철
• 소재민
• 윤지혜
• 정승희

Assignees

• 성균관대학교산학협력단

Dates

Publication Date

20260507

Application Date

20241025

Claims (7)

1. A biomarker composition for predicting the differentiation ability of stem cells into cardiomyocytes, comprising as an active ingredient a preparation for measuring the expression or activity level of TEK and SDR42E1 genes or proteins.
2. In paragraph 1, A biomarker composition for predicting the differentiation ability of stem cells into cardiomyocytes, characterized in that the above preparation comprises one or more selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the mRNA of TEK and SDR42E1 genes.
3. In paragraph 1, A biomarker composition for predicting the differentiation ability of stem cells into cardiomyocytes, characterized in that the above preparation is an antibody or aptamer that specifically binds to TEK and SDR42E1 proteins.
4. In paragraph 1, A biomarker composition for predicting the differentiation ability of stem cells into cardiomyocytes, wherein the above-mentioned stem cells are of human origin.
5. A kit for predicting the differentiation ability of stem cells into myocardial cells, comprising a composition of any one of claims 1 to 4.
6. 1) A step of measuring the expression or activity levels of TEK and SDR42E1 genes or proteins in stem cells; 2) a step of comparing the expression or activity level of the TEK and SDR42E1 genes or proteins with a control group; and 3) A step of determining that the differentiation ability of stem cells into cardiomyocytes is reduced if the expression or activity level of the TEK and SDR42E1 genes or proteins is higher than that of the control group; A method for providing information for predicting the differentiation ability of stem cells into cardiomyocytes, comprising
7. 1) A step of treating stem cells with a candidate substance; 2) a step of measuring the expression or activity level of the TEK and SDR42E1 genes or proteins in the stem cells; and 3) a step of determining the candidate substance as a stem cell cardiomyocyte differentiation promoter if the expression or activity level of the TEK and SDR42E1 genes or proteins is reduced compared to before treatment with the candidate substance; comprising a screening method for stem cell cardiomyocyte differentiation promoters.

Description

Biomarker composition for predicting stem cell-derived cardiomyocyte differentiation potential containing TEK and SDR42E1 genes and method for predicting stem cell-derived cardiomyocyte differentiation potential through measurement of TEK and SDR42E1 The present invention relates to a biomarker composition for selecting induced pluripotent stem cell clones with high differentiation ability into stem cell-derived cardiomyocytes by detecting the expression of TEK and SDR42E1 genes. Specifically, it relates to a novel type of biomarker and a method for manufacturing the same, which allows for the efficient utilization of induced pluripotent stem cells and enables the evaluation of the stability of stem cell lines in clinical trials using pluripotent stem cell-derived cardiomyocytes by evaluating the stability of the differentiation ability of induced pluripotent stem cell clones using the expression levels of TEK and SDR42E1 genes as indicators. Human induced pluripotent stem cells (hiPSCs) possess self-renewal and pluripotency, making them useful in various fields and widely utilized as in vitro models for rare diseases and cell therapy. However, gene expression patterns differ between individuals or among stem cell lines generated from a single person, which significantly impacts differentiation potential. It is necessary to predict the stability of fabricated polyclonal induced pluripotent stem cells by analyzing the correlation between these gene expression patterns and differences in differentiation potential. Various analyses are conducted to evaluate fabricated induced pluripotent stem cells. To assess whether they possess the stem cell characteristics of pluripotency and self-renewal, established pluripotency marker expression and teratoma assays are performed. Even if these evaluation indicators are passed, stem cell lines with low differentiation potential and clonal heterogeneity are still being identified. Furthermore, stem cells differentiate into specific cell types by regulating their signaling pathways, and processes for regulating these pathways are employed to enhance efficiency. For example, in the case of myocardial differentiation using stem cells, the Wnt signaling pathway is regulated. Since the concentration of drugs regulating the signaling pathway varies depending on the stem cell line, this necessitates the laborious task of determining the optimal drug concentration. Additionally, in the case of myocardial differentiation, differentiation results can only be evaluated after a certain timeframe has passed according to the established differentiation protocol, resulting in significant time and cost consumption to obtain differentiation products for each stem cell line. As such, even among induced pluripotent stem cells whose stability has been evaluated using conventional technology, cases of differentiation potential vary; in particular, clonal heterogeneity occurs even in polyclones generated from a single individual. Therefore, this paper suggests the need for additional indicators to evaluate the stability of established induced pluripotent stem cells. Furthermore, it is necessary to devise a method to save time and costs by predicting the differentiation potential of induced pluripotent stem cells prior to the cardiomyocyte differentiation process. Against this backdrop, the inventors confirmed that the TEK and SDR42E1 genes are expressed in induced pluripotent stem cells that have a low degree of differentiation from cardiomyocytes (CM), i.e., have instability. They confirmed that the stability of polyclonal cells can be predicted through the difference in expression of these two genes (TEK and SDR42E1), and confirmed the potential for their use as biomarkers to predict the differentiation ability of induced pluripotent stem cells. Figure 1 shows the results of analyzing the generation and characteristics of human induced pluripotent stem cell (hiPSC) clones. Figure 2 shows the results of evaluating the differentiation efficiency and function of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). Figure 3 shows the results of confirming the phosphorylation status of the ERK1/2 signaling pathway increased in non-productive clones (NPCs). Figure 4 shows the transcriptome analysis results of human induced pluripotent stem cell (hiPSC) clones using bulk RNA sequencing. Figure 5 is a graph evaluating the change in expression levels of endogenous retroelement (ERE). Figure 6 is a graph showing the various chromatin states of human induced pluripotent stem cell (hiPSC) clones using ATAC sequencing. Figure 7 is a graph confirming that TEK and SDR42E1 are suitable marker genes for predicting cardiac differentiation ability. The present invention will be described below with reference to the attached drawings. However, the present invention may be embodied in various different forms and is therefore not limited to the embodiments described herein. When a part is describ