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Generation of Induced Pluripotent Stem Cells from Human Melanoma Tumor-infiltrating Lymphocytes
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MiR-302-Mediated Somatic Cell Reprogramming and Method for Generating Tumor-Free iPS Cells Using miR-302.
Shi-Lung Lin1, Jack S Chen1, Shao-Yao Ying2
1WJWU & LYNN Institute for Stem Cell Research, Santa Fe Springs, CA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2020
Summary
Human induced pluripotent stem cells (iPSCs) can form tumors, but miR-302 microRNA prevents this by regulating cell cycle and tumor suppressor genes, creating safer iPSCs for therapy.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Human induced pluripotent stem cells (iPSCs) carry risks of teratoma formation and tumorigenicity.
- Naturally occurring embryonic stem cells (ESCs) possess self-renewal and pluripotency without tumor risk.
- Tumor prevention mechanisms in ESCs remain largely uncharacterized.
Purpose of the Study:
- To investigate the mechanism of tumor suppression in ESCs.
- To identify factors regulating iPSC tumorigenicity.
- To develop tumor-free iPSCs for therapeutic applications.
Main Methods:
- Examined cell cycle gene regulation in embryonic cells.
- Investigated the role of ESC-specific microRNA (miRNA), miR-302.
- Analyzed miR-302's effect on cell cycle pathways (cyclin E-CDK2, cyclin D-CDK4/6) and tumor suppressor genes (p16Ink4a, p14/p19Arf).
Main Results:
- miR-302 co-suppresses cell cycle pathways (G1-S transition) and silences BMI-1, a cancer stem cell marker.
- miR-302 promotes expression of tumor suppressor genes p16Ink4a and p14/p19Arf.
- This leads to attenuated cell cycle rates in iPSCs, similar to early embryonic cells, preventing tumor formation.
Conclusions:
- miR-302 effectively suppresses iPSC tumorigenicity by regulating cell cycle and activating tumor suppressors.
- miR-302 also initiates DNA methylation, activates ESC-specific genes, and inhibits developmental signaling.
- A protocol to express miR-302 naturally generates tumor-free iPSCs for safe biological and therapeutic use.
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