Genomic Safe Harbor Expression of PAX7 for the Generation of Engraftable Myogenic Progenitors

Kim, Hyunkee; Selvaraj, Sridhar; Kiley, James P.; Azzag, Karim; Garay, Bayardo I.; Perlingeiro, Rita C.R.

doi:10.1016/j.stemcr.2020.11.001

Cited by 23 publications

(21 citation statements)

References 35 publications

(43 reference statements)

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“…As anticipated, the vast majority of the Pax7 + cell fraction was from the host and therefore negative for LMNA ( Figures 4D and 4E ). This level of engraftment is consistent with human-to-mouse transplantation ( Kim et al, 2021 ), and the use of non-irradiated muscles, in which the recipient’s satellite cell compartment is preserved.…”

Section: Resultssupporting

confidence: 81%

A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy

et al. 2021

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Section: Resultssupporting

confidence: 81%

A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy

et al. 2021

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“…Although transgene delivery through viral vectors may raise concern of insertional mutagenesis, the development of safe self-inactivating lentiviral vectors [ 93 ], which have resulted in successful lentiviral-based gene therapy clinical trials [ 94 ] provide evidence of safety. Alternatively, the AAVS1 genomic safe harbor locus can be used for genetic modification [ 87 ].…”

Section: Discussionmentioning

confidence: 99%

“…The use of small molecules enhances the efficiency of PAX7-induced myogenic specification and reduces the variability among independent PSC lines, but these do not replace PAX7 induction [ 85 ]. In a recent study, Kim and colleagues utilized a doxycycline-inducible PAX7 transgene inserted into the genomic safe harbor locus, AAVS1 [ 86 ], for the alternative strategy of lentiviral PAX7 delivery to generate PAX7 + myogenic progenitors [ 87 ]. This strategy showed regenerating muscle fibers upon the transplantation into CTX-injured TA muscles of NSG, NSG-mdx4 Cv , and C3KO-NSG [ 87 ].…”

Section: Differentiation Of Human Pscs Into Myogenic Progenitorsmentioning

confidence: 99%

“…In a recent study, Kim and colleagues utilized a doxycycline-inducible PAX7 transgene inserted into the genomic safe harbor locus, AAVS1 [ 86 ], for the alternative strategy of lentiviral PAX7 delivery to generate PAX7 + myogenic progenitors [ 87 ]. This strategy showed regenerating muscle fibers upon the transplantation into CTX-injured TA muscles of NSG, NSG-mdx4 Cv , and C3KO-NSG [ 87 ]. Other groups have used PAX7 induction to generate human skeletal muscle from PSCs [ 88 , 89 ], notably Rao and colleagues generated skeletal muscle bundles, which upon implantation into muscles of NSG or nude mice, survived, vascularized and maintained functionality [ 89 ].…”

Section: Differentiation Of Human Pscs Into Myogenic Progenitorsmentioning

confidence: 99%

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Generation of human myogenic progenitors from pluripotent stem cells for in vivo regeneration

Kim

Perlingeiro

2022

Cell. Mol. Life Sci.

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Muscular dystrophy encompasses a large number of heterogeneous genetic disorders characterized by progressive and devastating muscle wasting. Cell-based replacement strategies aimed at promoting skeletal muscle regeneration represent a candidate therapeutic approach to treat muscular dystrophies. Due to the difficulties of obtaining large numbers of stem cells from a muscle biopsy as well as expanding these in vitro, pluripotent stem cells (PSCs) represent an attractive cell source for the generation of myogenic progenitors, given that PSCs can repeatedly produce large amounts of lineage-specific tissue, representing an unlimited source of cells for therapy. In this review, we focus on the progress to date on different methods for the generation of human PSC-derived myogenic progenitor cells, their regenerative capabilities upon transplantation, their potential for allogeneic and autologous transplantation, as well as the specific challenges to be considered for future therapeutic applications.

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“…Several protocols are currently available to generate skeletal myogenic derivatives from hiPSCs (reviewed in (Selvaraj, Kyba and Perlingeiro, 2019)). Starting from the pioneering studies based upon controlled expression of myogenic regulators to obtain transplantable skeletal myogenic cells from hiPSCs (e.g., Darabi et al, 2012;Goudenege et al, 2012;Tedesco et al, 2012), the field has refined transgene-based protocols to direct hiPSC differentiation into skeletal muscle (e.g., Albini et al, 2013;Maffioletti et al, 2015;Shoji et al, 2016;Kim et al, 2021), whilst also developing genomic-integration-free, small molecule-based methods to derive myogenic cells mimicking embryonic development (e.g., Borchin et al, 2013;Caron et al, 2016;Chal et al, 2016;Hicks et al, 2018). However, the focus on perfecting methods to obtain myogenic progenitors resembling self-renewing MuSCs has neglected the critical need to enhance their migration capacity, which is essential to deliver cells to large or multiple muscle districts.…”

Section: Introductionmentioning

confidence: 99%

DLL4 and PDGF-BB regulate migration of human iPSC-derived skeletal myogenic progenitors

Ferrari

Choi

Moyle

et al. 2021

Preprint

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Muscle satellite stem cells (MuSCs) are responsible for skeletal muscle growth and regeneration. Despite their differentiation potential, human MuSCs have limited in vitro amplification and in vivo migration capacity, limiting their use in cell therapies for diseases affecting multiple skeletal muscle groups such as muscular dystrophies. Several protocols have been established to derive progenitor cells similar to MuSCs from human induced pluripotent stem cells (hiPSCs), in order to establish a source of myogenic cells with controllable proliferation and differentiation capacity. However, currently available hiPSC myogenic derivatives also suffer from limitations of cell migration, ultimately delaying their clinical translation. Here we provide evidence that activation of NOTCH and PDGF pathways with DLL4 and PDGF-BB improves migration of hiPSC-derived myogenic progenitors in vitro. Transcriptomic and functional analyses demonstrate that this property is conserved across species and multiple hiPSC lines, including genetically-corrected hiPSC derivatives from a patient with Duchenne muscular dystrophy. DLL4 and PDGF-BB treatment had no negative impact on cell proliferation; cells maintained their myogenic memory, with differentiation fully rescued by NOTCH inhibition. RNAseq analysis indicate that pathways involved in cell migration are modulated in treated myogenic progenitors, consistent with results from functional profiling of cell motility at single cell resolution. Notably, treated cells also showed enhanced trans-endothelial migration in transwell assays. Enhancing extravasation is a key translational milestone for intravascular delivery of hiPSC myogenic derivatives: our study establishes the foundations of a transgene-free, developmentally inspired strategy to achieve this goal, moving hiPSCs one step closer to future muscle gene and cell therapies.

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Genomic Safe Harbor Expression of PAX7 for the Generation of Engraftable Myogenic Progenitors

Cited by 23 publications

References 35 publications

A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy

A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy

Generation of human myogenic progenitors from pluripotent stem cells for in vivo regeneration

DLL4 and PDGF-BB regulate migration of human iPSC-derived skeletal myogenic progenitors

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