Enhancing oligodendrocyte differentiation by transient transcription activation via DNA nanoparticle-mediated transfection.
Current approaches to derive oligodendrocytes from human pluripotent stem cells (hPSCs) need extended exposure of hPSCs to growth factors and small molecules, which limits their clinical application because of the lengthy culture time required and low generation efficiency of myelinating oligodendrocytes. Compared to extrinsic growth factors and molecules, oligodendrocyte differentiation and maturation can be more effectively modulated by regulation of the cell transcription network. In the developing central nervous system (CNS), two basic helix-loop-helix transcription factors, Olig1 and Olig2, are decisive in oligodendrocyte differentiation and maturation. Olig2 plays a critical role in the specification of oligodendrocytes and Olig1 is crucial in promoting oligodendrocyte maturation. Recently viral vectors have been used to overexpress Olig2 and Olig1 in neural stem/progenitor cells (NSCs) to induce the maturation of oligodendrocytes and enhance the remyelination activity in vivo. Because of the safety issues with viral vectors, including the insertional mutagenesis and potential tumor formation, non-viral transfection methods are preferred for clinical translation. Here we report a poly(β-amino ester) (PBAE)-based nanoparticle transfection method to deliver Olig1 and Olig2 into human fetal tissue-derived NSCs and demonstrate efficient oligodendrocyte differentiation following transgene expression of Olig1 and Olig2. This approach is potentially translatable for engineering stem cells to treat injured or diseased CNS tissues.



Current protocols to derive oligodendrocytes from human pluripotent stem cells (hPSCs) require lengthy culture time with low generation efficiencies of mature oligodendrocytes. We described a new approach to enhance oligodendrocyte differentiation through nanoparticle-mediated transcription modulation. We tested an effective transfection method using cell-compatible poly (β-amino ester) (PBAE)/DNA nanoparticles as gene carrier to deliver transcription factor Olig1 and Olig2 into human fetal tissue-derived neural stem/progenitor cells, and showed efficient oligodendrocyte differentiation following transgene expression of Olig1 and Olig2. We believe that this translatable approach can be applied to many other cell-based regenerative therapies as well.
Translational Tissue Engineering Center, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Department of Materials Science & Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.
Xiaowei
X
Li
Translational Tissue Engineering Center, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Stephany Y
SY
Tzeng
Translational Tissue Engineering Center, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Camila Gadens
CG
Zamboni
Department of Pathology, Division of Neuropathology, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Department of Neurology, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA; Department of Psychiatry & Behavioral Sciences, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA.
Vassilis E
VE
Koliatsos
Department of Neurology, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA; Department of Psychiatry & Behavioral Sciences, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Institute for Cell Engineering, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA; The Solomon H. Snyder Department of Neuroscience, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Guo-Li
GL
Ming
Translational Tissue Engineering Center, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Department of Materials Science & Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Jordan J
JJ
Green
Translational Tissue Engineering Center, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA; Department of Materials Science & Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA. Electronic address: hmao@jhu.edu.
Hai-Quan
HQ
Mao
D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics; D000076104:Oligodendrocyte Transcription Factor 2 / Q000096:biosynthesis / Q000235:genetics; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics; D014162:Transfection / Q000379:methods*
D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; D000076104:Oligodendrocyte Transcription Factor 2; C405707:OLIG1 protein, human; C405708:OLIG2 protein, human
Nanoparticles; Olig1; Olig2; Oligodendrocyte differentiation; Transfection