Ips cell-derived cardiogenicity is hindered by sustained integration of reprogramming transgenes

Almudena Martinez-Fernandez, Timothy J Nelson, Santiago Reyes, Alexey E. Alekseev, Frank Secreto, Carmen M Terzic, Rosanna Beraldi, Hoon Ki Sung, Andras Nagy, Andre Terzic

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background: Nuclear reprogramming inculcates pluripotent capacity by which de novo tissue differentiation is enabled. Yet, introduction of ectopic reprogramming factors may desynchronize natural developmental schedules. This study aims to evaluate the effect of imposed transgene load on the cardiogenic competency of induced pluripotent stem (iPS) cells.Methods and Results: Targeted inclusion and exclusion of reprogramming transgenes (c-MYC, KLF4, OCT4, and SOX2) was achieved using a drug-inducible and removable cassette according to the piggyBac transposon/transposase system. Pulsed transgene overexpression, before iPS cell differentiation, hindered cardiogenic outcomes. Delayed in counterparts with maintained integrated transgenes, transgene removal enabled proficient differentiation of iPS cells into functional cardiac tissue. Transgene-free iPS cells generated reproducible beating activity with robust expression of cardiac a-actinin, connexin 43, myosin light chain 2a, a/ß-myosin heavy chain, and troponin I. Although operational excitation-contraction coupling was demonstrable in the presence or absence of transgenes, factor-free derivatives exhibited an expedited maturing phenotype with canonical responsiveness to adrenergic stimulation.Conclusions: A disproportionate stemness load, caused by integrated transgenes, affects the cardiogenic competency of iPS cells. Offload of transgenes in engineered iPS cells ensures integrity of cardiac developmental programs, underscoring the value of nonintegrative nuclear reprogramming for derivation of competent cardiogenic regenerative biologics.

Original languageEnglish (US)
Pages (from-to)667-676
Number of pages10
JournalCirculation: Cardiovascular Genetics
Volume7
Issue number5
DOIs
StatePublished - Oct 1 2014

Fingerprint

Transgenes
Induced Pluripotent Stem Cells
Transposases
Actinin
Excitation Contraction Coupling
Myosin Light Chains
Connexin 43
Troponin I
Myosin Heavy Chains
Biological Products
Adrenergic Agents
Cell Differentiation
Appointments and Schedules
Phenotype
Pharmaceutical Preparations

Keywords

  • Induced pluripotent stem cells
  • Nuclear reprogramming
  • Regeneration
  • Regenerative medicine
  • Stem cells

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Genetics(clinical)
  • Genetics

Cite this

Ips cell-derived cardiogenicity is hindered by sustained integration of reprogramming transgenes. / Martinez-Fernandez, Almudena; Nelson, Timothy J; Reyes, Santiago; Alekseev, Alexey E.; Secreto, Frank; Terzic, Carmen M; Beraldi, Rosanna; Sung, Hoon Ki; Nagy, Andras; Terzic, Andre.

In: Circulation: Cardiovascular Genetics, Vol. 7, No. 5, 01.10.2014, p. 667-676.

Research output: Contribution to journalArticle

Martinez-Fernandez, Almudena ; Nelson, Timothy J ; Reyes, Santiago ; Alekseev, Alexey E. ; Secreto, Frank ; Terzic, Carmen M ; Beraldi, Rosanna ; Sung, Hoon Ki ; Nagy, Andras ; Terzic, Andre. / Ips cell-derived cardiogenicity is hindered by sustained integration of reprogramming transgenes. In: Circulation: Cardiovascular Genetics. 2014 ; Vol. 7, No. 5. pp. 667-676.
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AU - Reyes, Santiago

AU - Alekseev, Alexey E.

AU - Secreto, Frank

AU - Terzic, Carmen M

AU - Beraldi, Rosanna

AU - Sung, Hoon Ki

AU - Nagy, Andras

AU - Terzic, Andre

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