Optimizing Therapeutic Revascularization by Endothelial Cell Transplantation

通过内皮细胞移植优化治疗性血运重建

• 批准号: 9516109
• 负责人: JORDAN S POBER
• 金额: $ 37.24万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-26 至 2019-09-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9516109
• 关键词: 3D Print; Address; Adult; Allogenic; Animal Sources; Animals; Apligraf; Attention; Autologous; Basement membrane; Beds; Biological; Blood Vessels; Cattle; Cell Line; Cell Transplantation; Cells; Chronic; Clinical; Clinical Trials; Collaborations; Collagen; Collagen Type I; Collagen Type IV; Culture Media; Cutaneous; Dermal; Dermis; Development; Elastin; Elderly; Eligibility Determination; Endothelial Cells; Engineering; Engraftment; Epidermis; Evaluation; Exposure to; Extracellular Matrix; FDA approved; Failure; Fibroblasts; Fibronectins; Funding; Generations; Goals; Grant; Harvest; Histology; Human; Human Engineering; Hyaluronic Acid; Immune system; Immunodeficient Mouse; Impaired wound healing; Impairment; Implant; In Vitro; Individual; Infection; Ink; Institutes; Jordan; Karyotype; Laminin; Medical; Membrane Proteins; Morbidity - disease rate; Morphology; Mus; Muscle Cells; Natural Products; Neonatal; Newborn Infant; Papillary; Parents; Perfusion; Pericytes; Preparation; Primary Lesion; Printing; Production; Proteins; Recombinants; Regenerative Medicine; Research Project Grants; Rheology; Risk; Sirolimus; Site; Skin; Skin Substitutes; Skin Ulcer; Skin graft; Source; System; Therapeutic; Thick; Tissue Engineering; Tissues; Transmission Electron Microscopy; Ulcer; Umbilical Cord Blood; Vascular Smooth Muscle; Vascular blood supply; Work; adult stem cell; angiogenesis; biophysical techniques; bioprinting; cell type; clinical development; clinical practice; curative treatments; diabetic; experimental study; fetal bovine serum; healing; human tissue; immunocytochemistry; innovation; interstitial; keratinocyte; macromolecule; method development; novel strategies; paracrine; parent grant; peripheral blood; pre-clinical; response; self assembly; senescence; success; vascular tissue engineering

7. PROJECT SUMMARY/ABSTRACT The overall goal of this research project is to develop a vascularized full thickness skin substitute, engraftment of which can provide a definitive clinical cure of chronic cutaneous ulcers, an unmet medical need. This application is a competitive Revision to add a specific aim to a funded R01 grant (HL-085416 “Optimizing Therapeutic Revascularization by Endothelial Cell Transplantation”) submitted in response to RFA-HL-17-029 (Revision Applications for Regenerative Medicine Innovation Projects). The parent R01 grant is focused on establishing perfusion of tissue engineered grafts through the self-assembly of incorporated human endothelial cells (EC) into microvessels. We have found that the best source of EC for this purpose is the differentiated progeny of human endothelial colony forming cells (HECFC), an “adult” stem cell type isolated from neonatal cord or adult peripheral blood as defined by the terms of this RFA. Over the past six months, Drs. W. Mark Saltzman and Jordan S. Pober, co-PIs of the parent grant, have established a collaboration with Dr. Pankaj Karande to apply their approach for microvessel generation to his approach for 3D printing to create a full thickness human skin substitute. Current FDA-approved skin substitutes have all had only limited clinical success because they fail to vascularize, and consequently slough over the course of a few weeks. The vascularized skin substitute produced through this collaboration can potentially address this problem. However, the skin substitutes that we have generated to date employ bio-inks containing human cells that have been cultured in fetal bovine serum and use extracellular matrix molecules derived from animal sources. Similar exposures to animal proteins used in the production of currently approved skin substitutes have further limited their approved use. The goal of our new specific aim is to develop a vascularized full thickness human skin substitute through 3D printing made with bio-inks that are composed of human cells that have not been exposed to animal proteins and extracellular matrix molecules that are either derived from human tissues or made by recombinant approaches that do not involve exposure to animal proteins. Subaims a and b describe our approaches to achieve this. Subaim c will involve evaluation of perfusion of the resultant 3D bioprinted skin graft compared to natural skin and to Apligraf, a currently approved full thickness skin substitute, after engraftment on immunodeficient mouse hosts. The development of methods for creating a fully humanized and vascularized full thickness skin equivalent will provide the information needed to develop Good Manufacturing Practice (GMP) conditions for preparing clinical grade skin substitutes.

7.项目总结/摘要 本研究项目的总体目标是开发一种血管化的全层皮肤替代物， 其可以提供慢性皮肤溃疡的确定的临床治愈，这是一种未满足的医疗需求。这 申请是一个竞争性的修订，以增加一个具体的目标，以资助R 01赠款（HL-085416“优化 通过内皮细胞移植进行的治疗性血运重建”）提交，以回复RFA-HL-17-029 （再生医学创新项目的修订申请）。父R 01补助金的重点是 通过掺入的人内皮细胞的自组装建立组织工程移植物的灌注 细胞（EC）进入微血管。我们发现，用于此目的的EC的最佳来源是分化的 人内皮集落形成细胞（HECFC）的后代，一种从新生儿分离的“成人”干细胞类型， 本RFA术语定义的脐带血或成人外周血。在过去的六个月里，W.马克 Saltzman和Jordan S. Pober，父母补助金的共同PI，已经与Pankaj博士建立了合作关系。 Karande将他们的微血管生成方法应用于他的3D打印方法，以创建一个完整的 厚的人类皮肤替代品。目前FDA批准的皮肤替代品都只有有限的临床应用， 成功是因为它们不能血管化，因此在几周内斯劳。的 通过这种合作生产的血管化皮肤替代品可能解决这个问题。 然而，我们迄今为止生产的皮肤替代品采用含有人类细胞的生物墨水， 已经在胎牛血清中培养并使用来源于动物来源的细胞外基质分子。 在目前批准的皮肤替代品的生产中使用的动物蛋白的类似暴露， 限制其使用。我们新的具体目标的目标是开发一个血管化的全层人类 皮肤替代品通过3D打印制成，由人体细胞组成， 暴露于动物蛋白质和细胞外基质分子，所述细胞外基质分子来源于人组织， 通过不涉及暴露于动物蛋白的重组方法制造。子项a和B描述了 我们的方法来实现这一点。Subaim c将涉及对所得3D生物打印皮肤的灌注的评估。 移植相比，天然皮肤和Aplimetry，目前批准的全厚度皮肤替代品， 在免疫缺陷小鼠宿主上的移植。制定方法，创造一个完全人性化的， 血管化全层皮肤等同物将提供开发良好制造所需的信息 制备临床级皮肤替代品的GMP条件。