Inducible Forward Programming of Human Pluripotent Stem Cells to Hemato-endothelial Progenitor Cells with Hematopoietic Progenitor Potential

Lange, Lucas; Hoffmann, D.; Schwarzer, Adrian; Ha, Teng-Cheong; Philipp, Friederike; Lenz, Daniela; Morgan, Michael M.; Schambach, Axel

doi:10.1016/j.stemcr.2020.05.019

Cited by 13 publications

(21 citation statements)

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“…A recently described inducible transcription factor-mediated forward programming approach to efficiently produce large numbers of hemato-endothelial progenitor cells and hematopoietic progenitor cells may also become useful for generating "off-the-shelf " cell therapies, such as CAR NK cells (57). While this strategy led to sustained production of myeloid lineages, differentiation into the lymphoid lineages was less robust.…”

Section: Ipsc and Other Cell Sourcesmentioning

confidence: 99%

Use of Cell and Genome Modification Technologies to Generate Improved “Off-the-Shelf” CAR T and CAR NK Cells

et al. 2020

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The broad success of adoptive immunotherapy to treat human cancer has resulted in a paradigm shift in modern medicine. Modification of autologous and allogenic immune cells with chimeric antigen receptors (CAR) designed to target specific antigens on tumor cells has led to production of CAR T and CAR NK cell therapies, which are ever more commonly introduced into cancer patient treatment protocols. While allogenic T cells may offer advantages such as improved anti-tumor activity, they also carry the risk of adverse reactions like graft-versus-host disease. This risk can be mitigated by use of autologous immune cells, however, the time needed for T and/or NK cell isolation, modification and expansion may be too long for some patients. Thus, there is an urgent need for strategies to robustly produce "off-the-shelf" CAR T and CAR NK cells, which could be used as a bridging therapy between cancer diagnosis or relapse and allogeneic transplantation. Advances in genome modification technologies have accelerated the generation of designer cell therapy products, including development of "off-the-shelf" CAR T cells for cancer immunotherapy. The feasibility and safety of such approaches is currently tested in clinical trials. This review will describe cell sources for CAR-based therapies, provide background of current genome editing techniques and the applicability of these approaches for generation of universal "off-the-shelf" CAR T and NK cell therapeutics.

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Section: Ipsc and Other Cell Sourcesmentioning

confidence: 99%

Use of Cell and Genome Modification Technologies to Generate Improved “Off-the-Shelf” CAR T and CAR NK Cells

et al. 2020

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“…The iPSC differentiation in hematopoietic progenitors is based on the forward programming protocol. 30 iPSC models were transduced with pRRL.PPT.T11.hSCL.2A .hLMO2.PGK.M2.2A.Puro.pre (MOI 1) and pRRL.PPT .T11.GATA2.2A.ETV2.PGK.M2.2A.Zeo.pre (MOI 10). Positive selection based on antibiotics was performed.…”

Section: Generation Of Macrophages By Forward Programmingmentioning

confidence: 99%

“…Positive selection based on antibiotics was performed. Positively transduced cells were differentiated according to the forward programming protocol described by Lange et al 30 For macrophage differentiation, the protocol was changed and from day 11 to 15, cells were cultured in X-VIVO 15 (Biozym Scientific GmbH, Hessisch Oldendorf, Germany) medium supplemented with 100 U/mL IL-10-ASSOCIATED KNOCKOUT IPSCS FOR DISEASE MODELING penicillin, 100 lg/mL streptomycin, 100 ng/mL SCF, 50 ng/mL macrophage colony-stimulating factor (M-CSF), 10 ng/mL IL-6, and 25 ng/mL IL-3 for myeloid/monocyte specification. From day 15 to 22, cells were cultured in Roswell Park Memorial Institute 1640 (RPMI 1640; PAN-Biotech, Aidenbach, Germany) medium supplemented with 100 U/mL penicillin, 100 lg/mL streptomycin, 10% heatinactivated fetal bovine serum (FBS), and 100 ng/mL M-CSF for terminal maturation into macrophages.…”

Section: Generation Of Macrophages By Forward Programmingmentioning

confidence: 99%

“…To produce large amounts of hematopoietic cells, a forward programming system based on inducible and regulated overexpression of the transcriptional regulators SCL, LMO2, GATA2, and ETV2 was used. 30 The expression of these transcriptional regulators strongly improves the generation of hemato-endothelial progenitor cells (HEPs) by gradual hemato-endothelial differentiation and HPC through endothelial-to-hematopoietic transition. The established KO iPSCs were transduced with the LVs of the forward programming system and during hemato-endothelial differentiation from day 0 to 7 (Phase I), HEPs (CD73 -/CD144 + ) were produced (Fig.…”

Section: Differentiation Of Ipsc Disease Models Into Functional Macrophagesmentioning

confidence: 99%

“…In addition to the differentiation of iPSCs via embryoid body (EB) formation and the myeloid cell forming complex (MCFC) to generate monocytes and macrophages, 29 a new forward programming protocol to produce hematopoietic progenitor cells (HPCs) through endothelial-to-hematopoietic transition is a promising approach to generate disease models. 30 In the present study, we aimed to further improve disease modeling for PIDs, to characterize the IL-10 signaling pathway and associated diseases, as well as to investigate potential treatment options by using an RNA-guided CRISPR-Cas9 system to introduce knockouts (KOs) in the IL-10 signaling pathway-associated IL-10RA and IL-10RB chains as well as in the downstream targets STAT1 and STAT3. To analyze the different IL-10 signaling defects, the iPSC KO models and a patient-derived disease model were differentiated into macrophages.…”

Section: Introductionmentioning

confidence: 99%

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Knockout-Induced Pluripotent Stem Cells for Disease and Therapy Modeling of IL-10-Associated Primary Immunodeficiencies

Sens¹,

Hoffmann²,

Lange³

et al. 2021

Human Gene Therapy

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Samples from patients with rare diseases, such as primary immunodeficiencies, are often limited, which hampers careful analysis of the pathomechanisms involved in immune cell dysregulation. To overcome this issue, induced pluripotent stem cells (iPSCs) represent an almost inexhaustible cell source and thus provide an excellent opportunity to generate disease models for rare diseases and to validate new therapeutic approaches. To obtain a better understanding of primary immunodeficiencies associated with the interleukin (IL)-10 signaling pathway, for example, very-early-onset inflammatory bowel disease (VEO-IBD), we generated genetic knockouts (KOs) of IL-10RA (IL-10 receptor a-chain) and IL-10RB (IL-10 receptor b-chain) as well as the downstream targets of the IL-10-receptor (IL-10R) signal transducers and activators of transcription (STAT)1 and STAT3 via an sgRNA (single-guide RNA)-CRISPR-Cas9-expressing lentiviral system. IL-10 signaling-associated KO models and a VEO-IBD patient-derived iPSC clone were differentiated into macrophages for disease models. IL-10R-or STAT3-deficient disease models showed no IL-10-induced BCL3 or SOCS3 expression, whereas lipopolysaccharide (LPS) stimulation induced IL-10R independently of BCL3 and SOCS3 expression. Cytokine secretion profiles from iPSC-derived macrophage disease models showed that IL-10 was involved in many inflammatory cytokine secretions, which indicated formation of both anti-and proinflammatory macrophage phenotypes. Macrophage-secreted cytokines were separated into IL-10R-and STAT3-dependent (IL-6, TNF-a), or into IL-10R-, STAT1-, and STAT3-dependent cytokines (CCL2, CXCL10). Importantly, lentiviral correction restored IL-10mediated regulation of LPS-induced cytokine secretion in corrected IL-10RB, STAT1, and VEO-IBD patient-derived disease models. Furthermore, treatment of IL-10RB-deficient macrophages with anti-inflammatory small molecules (SB202190, filgotinib) reduced proinflammatory cytokine secretion patterns. Taken together, the described iPSC KO models gave new insights into the pathomechanisms of immune cell dysregulation and served as model systems to test potential therapeutic approaches, including lentiviral gene therapy and targeted small-molecule treatment.

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Progress in the treatment of neonatal hypoxic-ischemic encephalopathy with umbilical cord blood mononuclear cells

Zhou,

Gao,

Tang

et al. 2023

Brain and Development

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Inducible Forward Programming of Human Pluripotent Stem Cells to Hemato-endothelial Progenitor Cells with Hematopoietic Progenitor Potential

Cited by 13 publications

References 0 publications

Use of Cell and Genome Modification Technologies to Generate Improved “Off-the-Shelf” CAR T and CAR NK Cells

Use of Cell and Genome Modification Technologies to Generate Improved “Off-the-Shelf” CAR T and CAR NK Cells

Knockout-Induced Pluripotent Stem Cells for Disease and Therapy Modeling of IL-10-Associated Primary Immunodeficiencies

Progress in the treatment of neonatal hypoxic-ischemic encephalopathy with umbilical cord blood mononuclear cells

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