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A synthetic mRNA cell reprogramming method using CYCLIN D1 promotes DNA repair generating improved genetically stable human induced pluripotent stem cells

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Autores de CIPF

Participantes ajenos a CIPF

  • Alvarez-Palomo, AB
  • Requena-Osete, J
  • Delgado-Morales, R
  • Grau-Bove, C
  • Tejera, AM
  • Otero, MJ
  • Barrot, C
  • Santos-Barriopedro, I
  • Vaquero, A
  • Mezquita-Pla, J
  • Moran, S
  • Naya, CH
  • Garcia-Martinez, I
  • Perez, FV
  • Blasco, MA
  • Esteller, M
  • Edel, MJ

Grupos de Investigación

Abstract

A key challenge for clinical application of induced pluripotent stem cells (iPSC) to accurately model and treat human pathologies depends on developing a method to generate genetically stable cells to reduce long-term risks of cell transplant therapy. Here, we hypothesized that CYCLIN D1 repairs DNA by highly efficient homologous recombination (HR) during reprogramming to iPSC that reduces genetic instability and threat of neoplastic growth. We adopted a synthetic mRNA transfection method using clinically compatible conditions with CYCLIN D1 plus base factors (OCT3/4, SOX2, KLF4, LIN28) and compared with methods that use C-MYC. We demonstrate that CYCLIN D1 made iPSC have (a) lower multitelomeric signal, (b) reduced double-strand DNA breaks, (c) correct nuclear localization of RAD51 protein expression, and (d) reduced single-nucleotide polymorphism (SNP) changes per chromosome, compared with the classical reprogramming method using C-MYC. CYCLIN D1 iPSC have reduced teratoma Ki67 cell growth kinetics and derived neural stem cells successfully engraft in a hostile spinal cord injury (SCI) microenvironment with efficient survival, differentiation. We demonstrate that CYCLIN D1 promotes double-stranded DNA damage repair predominantly through HR during cell reprogramming to efficiently produce iPSC. CYCLIN D1 reduces general cell stress associated with significantly lower SIRT1 gene expression and can rescue Sirt1 null mouse cell reprogramming. In conclusion, we show synthetic mRNA transfection of CYCLIN D1 repairs DNA during reprogramming resulting in significantly improved genetically stable footprint in human iPSC, enabling a new cell reprogramming method for more accurate and reliable generation of human iPSC for disease modeling and future clinical applications.

Datos de la publicación

ISSN/ISSNe:
1066-5099, 1549-4918

STEM CELLS  WILEY

Tipo:
Article
Páginas:
882-896
PubMed:
33621399

Citas Recibidas en Web of Science: 15

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Keywords

  • cellular therapy; induced pluripotent stem cells; neural stem cells (NSCs); cell cycle; clinical translation

Campos de Estudio

Proyectos asociados

Transplant of combined cell therapy form clinical grade iPSC-derived cells with neuroprotective small chemicals in a SCI rat model for central regeneration of spinal pathways

Investigador Principal: M VICTORIA MORENO MANZANO

FUNDACION LA MARATO DE TV3 . 2018

Combinatory treatment of Neural precursor cells and a new nanoconjugate of Fasudil for the clinical application in Acute Spinal Cord Injury

Investigador Principal: MARIA JESUS VICENT DOCON

FUNDACION LA MARATO DE TV3 . 2018

Nuevo biomaterial bio-activo para la regeneración de lesiones medulares

Investigador Principal: M VICTORIA MORENO MANZANO

MINISTERIO DE ECON. Y COMPET. . 2019

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