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.项目摘要/摘要 这项研究项目的总体目标是开发一种血管化的全层皮肤替代品，植入物 它可以为慢性皮肤溃疡提供明确的临床治疗方法，这是一种尚未得到满足的医疗需求。这 申请是一项竞争性修订，旨在为已资助的R01拨款(HL-085416“优化”)增加一个特定目标 通过内皮细胞移植进行治疗性血管重建术》) (修订再生医学创新项目申请书)。家长R01奖助金的重点是 通过自组装人内皮细胞建立组织工程移植物的灌流 细胞(EC)转化为微血管。我们发现，用于这一目的的EC的最佳来源是差异化的 人内皮细胞集落形成细胞(HECFC)的后代--一种从新生儿分离的成体干细胞类型 脐带血或成人外周血，如本RFA术语所定义。在过去的六个月里，W.Mark博士 索尔兹曼和乔丹·S·波伯，父母基金的联合私人投资者，已经与潘卡吉博士建立了合作关系 卡兰德将他们的微血管生成方法应用到他的3D打印方法中，以创建完整的 厚厚的人体皮肤替代品。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形成，结果在几周内就崩溃了。这个 通过这种合作生产的血管化皮肤替代品可以潜在地解决这个问题。 然而，到目前为止，我们已经产生的皮肤替代品使用的生物墨水含有人体细胞 已经在胎牛血清中培养，并使用来自动物来源的细胞外基质分子。 与目前批准的皮肤替代品生产中使用的动物蛋白的类似暴露进一步 限制了他们被批准的用途。我们新目标的具体目标是开发一个血管化的全身人 用生物墨水制成的3D打印皮肤替代品，这些墨水是由尚未被 接触动物蛋白和细胞外基质分子，这些分子来自人类组织或 通过不涉及接触动物蛋白的重组方法制成。苏巴姆a和b描述了 我们实现这一目标的方法。Subaim c将评估所得到的3D生物打印皮肤的灌注度。 移植皮肤与天然皮肤和目前批准的全层皮肤替代品Apligraf相比， 在免疫缺陷小鼠宿主上植入。为创造一个完全人性化和 血管化全厚皮肤等价物将提供发展良好制造所需的信息 生产临床级皮肤替代物的实践(GMP)条件。