Progenitor-derived human endothelial cells evade alloimmunity by CRISPR/Cas9-mediated complete ablation of MHC expression

Merola, Jonathan; Reschke, Melanie; Pierce, Richard W.; Qin, Lingfeng; Spindler, Susann; Baltazar, Tania; Manes, Thomas D.; López-Giráldez, Francesc; Li, Guangxin; Bracaglia, Laura G.; Xie, Catherine; Kirkiles-Smith, Nancy C.; Saltzman, W. Mark; Tietjen, Gregory T.; Tellides, George; Pober, Jordan S.

doi:10.1172/jci.insight.129739

Cited by 22 publications

(18 citation statements)

References 62 publications

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“…[61][62][63] While grafts containing unmodified human ECs will be alloimmunogenic, this property can be eliminated from HECFC-derived ECs by CRISPR/Cas9mediated deletion of HLA antigen expression. [64][65][66] We anticipate that successful engraftment of a 3D bioprinted human skin graft by incorporating allogeneic ECs without triggering rejection in the host would be of great clinical utility and the basis for the creation of an ''off-the-shelf'' clinical product.…”

Section: D Bioprinting Of Vascularized Human Skin Graftmentioning

confidence: 99%

Three Dimensional Bioprinting of a Vascularized and Perfusable Skin Graft Using Human Keratinocytes, Fibroblasts, Pericytes, and Endothelial Cells

Baltazar

Merola

Catarino

et al. 2020

Tissue Engineering Part A

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Multilayered skin substitutes comprising allogeneic cells have been tested for the treatment of nonhealing cutaneous ulcers. However, such nonnative skin grafts fail to permanently engraft because they lack dermal vascular networks important for integration with the host tissue. In this study, we describe the fabrication of an implantable multilayered vascularized bioengineered skin graft using 3D bioprinting. The graft is formed using one bioink containing human foreskin dermal fibroblasts (FBs), human endothelial cells (ECs) derived from cord blood human endothelial colony-forming cells (HECFCs), and human placental pericytes (PCs) suspended in rat tail type I collagen to form a dermis followed by printing with a second bioink containing human foreskin keratinocytes (KCs) to form an epidermis. In vitro, KCs replicate and mature to form a multilayered barrier, while the ECs and PCs self-assemble into interconnected microvascular networks. The PCs in the dermal bioink associate with EC-lined vascular structures and appear to improve KC maturation. When these 3D printed grafts are implanted on the dorsum of immunodeficient mice, the human EC-lined structures inosculate with mouse microvessels arising from the wound bed and become perfused within 4 weeks after implantation. The presence of PCs in the printed dermis enhances the invasion of the graft by host microvessels and the formation of an epidermal rete.

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Section: D Bioprinting Of Vascularized Human Skin Graftmentioning

confidence: 99%

Three Dimensional Bioprinting of a Vascularized and Perfusable Skin Graft Using Human Keratinocytes, Fibroblasts, Pericytes, and Endothelial Cells

Baltazar

Merola

Catarino

et al. 2020

Tissue Engineering Part A

Self Cite

190

197

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“…Fifth, the therapeutic use of UCB-derived ECFCs, which display higher proliferative potential (up to 100 population doublings), enhanced angiogenic activity and greater telomerase activity as compared to circulating ECFCs [ 22 , 134 ], is currently not feasible [ 33 ]. On the one hand, the heterologous injection of umbilical cord blood-derived ECFCs may lead to an alloimmune response, both in vitro and in vivo, due to the expression of the of class I and class II major histocompatibility complex (MHC) molecules, which are, respectively, recognized by CD8 + and CD4 + T cells [ 135 , 136 ]. On the other hand, processing and freezing a sufficient amount of umbilical cord blood-derived ECFCs for each individual at birth, so that these could be banked until required during adulthood, could be exceedingly expensive at this moment [ 33 ].…”

Section: Manipulation Of Pro-angiogenic Signaling Pathways To Imprmentioning

confidence: 99%

Therapeutic Potential of Endothelial Colony-Forming Cells in Ischemic Disease: Strategies to Improve their Regenerative Efficacy

Faris

Negri

Perna

et al. 2020

IJMS

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Cardiovascular disease (CVD) comprises a range of major clinical cardiac and circulatory diseases, which produce immense health and economic burdens worldwide. Currently, vascular regenerative surgery represents the most employed therapeutic option to treat ischemic disorders, even though not all the patients are amenable to surgical revascularization. Therefore, more efficient therapeutic approaches are urgently required to promote neovascularization. Therapeutic angiogenesis represents an emerging strategy that aims at reconstructing the damaged vascular network by stimulating local angiogenesis and/or promoting de novo blood vessel formation according to a process known as vasculogenesis. In turn, circulating endothelial colony-forming cells (ECFCs) represent truly endothelial precursors, which display high clonogenic potential and have the documented ability to originate de novo blood vessels in vivo. Therefore, ECFCs are regarded as the most promising cellular candidate to promote therapeutic angiogenesis in patients suffering from CVD. The current briefly summarizes the available information about the origin and characterization of ECFCs and then widely illustrates the preclinical studies that assessed their regenerative efficacy in a variety of ischemic disorders, including acute myocardial infarction, peripheral artery disease, ischemic brain disease, and retinopathy. Then, we describe the most common pharmacological, genetic, and epigenetic strategies employed to enhance the vasoreparative potential of autologous ECFCs by manipulating crucial pro-angiogenic signaling pathways, e.g., extracellular-signal regulated kinase/Akt, phosphoinositide 3-kinase, and Ca2+ signaling. We conclude by discussing the possibility of targeting circulating ECFCs to rescue their dysfunctional phenotype and promote neovascularization in the presence of CVD.

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“… 80 , 81 To reduce the requirement for fetal tissue, a recent model instead utilized neonatal thymus, which has produced human cells capable of rejecting skin xenografts. 82 …”

Section: Micementioning

confidence: 99%

Humanization of Immunodeficient Animals for the Modeling of Transplantation, Graft Versus Host Disease, and Regenerative Medicine

et al. 2020

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The humanization of animals is a powerful tool for the exploration of human disease pathogenesis in biomedical research, as well as for the development of therapeutic interventions with enhanced translational potential. Humanized models enable us to overcome biologic differences that exist between humans and other species, while giving us a platform to study human processes in vivo. To become humanized, an immune-deficient recipient is engrafted with cells, tissues, or organoids. The mouse is the most well studied of these hosts, with a variety of immunodeficient strains available for various specific uses. More recently, efforts have turned to the humanization of other animal species such as the rat, which offers some technical and immunologic advantages over mice. These advances, together with ongoing developments in the incorporation of human transgenes and additional mutations in humanized mouse models, have expanded our opportunities to replicate aspects of human allotransplantation and to assist in the development of immunotherapies. In this review, the immune and tissue humanization of various species is presented with an emphasis on their potential for use as models for allotransplantation, graft versus host disease, and regenerative medicine.

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Progenitor-derived human endothelial cells evade alloimmunity by CRISPR/Cas9-mediated complete ablation of MHC expression

Cited by 22 publications

References 62 publications

Three Dimensional Bioprinting of a Vascularized and Perfusable Skin Graft Using Human Keratinocytes, Fibroblasts, Pericytes, and Endothelial Cells

Three Dimensional Bioprinting of a Vascularized and Perfusable Skin Graft Using Human Keratinocytes, Fibroblasts, Pericytes, and Endothelial Cells

Therapeutic Potential of Endothelial Colony-Forming Cells in Ischemic Disease: Strategies to Improve their Regenerative Efficacy

Humanization of Immunodeficient Animals for the Modeling of Transplantation, Graft Versus Host Disease, and Regenerative Medicine

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