Isochoric Pressure Based Preservation of Ovarian Tissue

基于等容压的卵巢组织保存

• 批准号: 10392764
• 负责人: Bradley P Weegman
• 金额: $ 58.78万
• 依托单位: SYLVATICA BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-26 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392764
• 关键词: Address; American; American Society of Clinical Oncology; Area; Autoimmune Diseases; Autologous; Autologous Transplantation; Biomedical Research; Blood Vessels; Bone Marrow; Cancer Survivor; Cell Volumes; Chemicals; Chemotherapy and/or radiation; Child; Childhood; Climacteric; Clinic; Clinical; Competence; Complement; Complex; Continuance of life; Cryopreservation; Cryopreserved Tissue; Development; Devices; Disease; Dry Ice; Embryonic Development; Endocrine; Engineering; Equilibrium; Equipment; Evaluation; Failure; Family suidae; Feasibility Studies; Fertility; Fishes; Food; Formulation; Foundations; Freedom; Freezing; Germ Cells; Gold; Growth; Hematological Disease; High temperature of physical object; Hormones; Human; Ice; Immune; Individual; Infertility; Lead; Letters; Malignant Childhood Neoplasm; Malignant Neoplasms; Medicine; Metabolic Diseases; Methods; Modeling; Mus; National Institute of Child Health and Human Development; Oocytes; Organ; Organ Donor; Organ Preservation; Ovarian; Ovarian Tissue; Ovary; Patients; Perfusion; Periodicity; Phase; Preservation Technique; Primates; Production; Progress Reports; Protective Agents; Protocols documentation; Public Health; Rattus; Recovery; Reproductive Health; Reproductive Medicine; Reproductive Physiology; Research; Research Priority; Science; Sickle Cell; Sickle Cell Anemia; Small Business Innovation Research Grant; Societies; Stretching; Surgical Replantation; Survival Rate; Suspensions; System; Technology; Temperature; Testicular Tissue; Testis; Thermodynamics; Tissue Engineering; Tissue Preservation; Tissues; Translating; Transplantation; United States National Institutes of Health; Validation; Vascular Graft; Xenograft procedure; base; biomaterial compatibility; boys; cancer therapy; clinical application; clinical practice; clinically relevant; combinatorial; commercialization; cryogenics; design; experience; fertility preservation; girls; improved; in vivo; innovation; minimally invasive; next generation; nonhuman primate; novel; novel strategies; offspring; oncofertility; pediatric patients; phase 2 study; preconditioning; prepuberty; preservation; pressure; prototype; reproductive organ; restoration; scale up; service member; sperm cell; success; survivorship; tool; trauma care; young adult; young woman

ABSTRACT This project combines and further develops our ice-free isochoric vitrification platform, multi-step/multi-thermic machine perfusion, and next generation non-toxic cryopreservation cocktails to find a viable solution to the large public health need for fertility preservation in children and young adults after different diseases and disease treatments. Such technologies are urgently needed by 650,000 children and young adult cancer survivors in US, to help preserve fertility and restore endocrine function post toxic treatments. The NIH has recognized this as a top priority, the fourth of NICHDs Fertility and Infertility Branch’s SBIR priority research areas being focused on “Novel techniques for preservation of gametes and whole ovary and testes.” Using carefully optimized protocols and cryostasis cocktails, our team and others have successfully cryopreserved multiple tissues in an ice-free vitreous “glassy” or “amorphous” state, allowing for indefinite storage. Unfortunately, these advances in ice-free preservation have mostly not been successfully scaled beyond small-tissue and small-volume cell suspensions due to the high cryoprotectant chemical concentrations, and rapid cooling/warming rates necessary for current vitrification methods. However, our broader group together with Dr. Rubinsky at UC-Berkeley, have developed a scalable and biocompatible isochoric (constant volume) cryopreservation paradigm for completely ice-free vitrification. Building on the success of the Phase I feasibility study, this project will deliver: (i) an improved preservation and banking of ovarian tissues strips via ice-free equilibrium isochoric vitrification (compared to current gold-standard slow- freezing), (ii) comprehensive protocol for banking whole human ovaries with functional validation in human to mouse xeno-transplants and validation in non-human primates with embryo development and eventually healthy offspring, and (iii) clinically relevant prototype systems of isochoric vitrification, capable of banking whole ovaries (testes, or other similar size tissues) for years until re-plantation. In addition, this project provides tools, equipment, solutions and protocols with highly-translatable technology toward banking of whole vital organs, and other vascular grafts, addressing widespread needs for breakthroughs in biopreservation – from improved biospecimen preservation to vital organ banking and transplantation. To meet these objectives, in four specific aims, we (i) with the help of thermodynamic profiling, further optimize the composition of cryostasis solutions for isochoric vitrification with minimal pressures, (ii) refine the biocompatibility of the cocktails and protocols using ovarian tissue strips, with human-to-mouse xenotransplants and human blood vessels contractile functional assessment, (iii) scale to whole human ovary vitrification with multi-thermic machine perfusion and clinical grade isochoric devices, and as a capstone, (iv) cryobank whole non-human primate ovaries with ovarian tissue strips in vivo quality evaluation through minimally invasive autotransplantation, embryo development and eventual healthy offspring. Success of these novel approaches, individually or in combination, can enable further breakthroughs in oncofertility, biopreservation research, and clinical practice.

抽象的 该项目结合并进一步开发了我们的无冰等容玻璃化平台、多步/多温机 灌注和下一代无毒冷冻保存鸡尾酒，为广大公众健康找到可行的解决方案 儿童和青少年在不同疾病和疾病治疗后保留生育能力的需要。 美国 650,000 名儿童和年轻癌症幸存者迫切需要技术来帮助保持生育能力 NIH 已将其视为重中之重，是 NICHD 的第四个优先事项。 生育与不孕不育分会的 SBIR 优先研究领域专注于“保存精子的新技术” 配子以及整个卵巢和睾丸。” 使用精心优化的方案和冷冻鸡尾酒，我们的团队和其他人成功地冷冻保存 多个组织处于无冰的玻璃体“玻璃状”或“无定形”状态，可以无限期储存。 无冰保存方面的进展大多尚未成功扩展到小组织和小体积细胞之外 由于高冷冻保护剂化学浓度和快速冷却/升温速率所需的悬浮液 然而，我们更广泛的团队与加州大学伯克利分校的鲁宾斯基博士一起开发了当前的玻璃化方法。 用于完全无冰玻璃化的可扩展且生物相容的等容（恒定体积）冷冻保存范例。 在第一阶段可行性研究成功的基础上，该项目将实现：(i) 改善保护和 通过无冰平衡等容玻璃化冷冻保存卵巢组织条带（与当前的金标准慢速玻璃化相比） 冷冻），（ii）储存整个人类卵巢的综合方案，并在人与小鼠之间进行功能验证 在非人类灵长类动物中进行异种移植和验证，使其胚胎发育并最终产生健康的后代，以及 (iii) 临床相关的等容玻璃化原型系统，能够储存整个卵巢（测试或其他 类似大小的组织）多年，直到重新种植。此外，该项目还提供工具、设备、解决方案和 具有高度可转化技术的协议，用于存储整个重要器官和其他血管移植物，解决 生物保存领域广泛需要突破——从改进生物样本保存到重要器官 银行业务和移植。 为了实现这些目标，在四个具体目标中，我们 (i) 在热力学分析的帮助下，进一步优化 用于以最小压力进行等容玻璃化的低温冷冻溶液的组成，(ii) 改进生物相容性 使用卵巢组织条、人对小鼠异种移植和人类血管的鸡尾酒和方案 收缩功能评估，(iii) 使用多热机器灌注进行整个人类卵巢玻璃化冷冻的规模和 临床级等容装置，并作为顶点，(iv) 冷冻库带有卵巢组织的整个非人类灵长类动物卵巢 通过微创自体移植、胚胎发育和最终的体内质量评估 这些新方法的成功，无论是单独还是组合，都可以实现进一步的突破。 肿瘤生育力、生物保存研究和临床实践。