Ice-free cryopreservation of whole pediatric testes for autologous banking and replantation.

整个儿科睾丸的无冰冷冻保存用于自体储存和再植。

• 批准号: 9919065
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 30万
• 依托单位: TISSUE TESTING TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9919065
• 关键词: Achievement; Address; Adolescent; Adult; Age; AlamarBlue; Animals; Antioxidants; Aplastic Anemia; Apoptosis; Apoptotic; Applications Grants; Autoimmune Diseases; Autologous; Autologous Transplantation; Autopsy; Back; Benign; Biological Assay; Biopsy; Birth; Blood Vessel Tissue; Breeding; Cancer Patient; Cancer Survivor; Cell Survival; Cells; Characteristics; Child; Childhood; Chronic Childhood Arthritis; Clinic; Complex; Cooley' s anemia; Cryopreservation; Cryopreserved Cell; Cytotoxic Chemotherapy; Development; Diagnosis; Dimethyl Sulfoxide; Dissection; Dysmyelopoietic Syndromes; Equilibrium; Evaluation; Exposure to; Failure; Fanconi' s Anemia; Feasibility Studies; Female; Fertility; Fertilization; Fertilization in Vitro; Fibrosis; Formulation; Frequencies; Future; Genetic; Germ Cells; Germ Lines; Heart Valves; Heating; Hematological Disease; High Dose Chemotherapy; Histologic; Histology; Hormonal; Hormones; Human; Ice; Immune; In Vitro; Infertility; Injury; Interview; Klinefelter' s Syndrome; Lead; Letters; Libido; Licensing; Life; Macaca mulatta; Magnetism; Maintenance; Male Infertility; Malignant Neoplasms; Metals; Methods; Modeling; Nitrogen; Non-Malignant; Oocytes; Operative Surgical Procedures; Outcome; Parents; Patients; Phase; Pre-implantation Embryo Development; Pregnancy; Production; Propylene Glycols; Protocols documentation; Puberty; Public Health; Radiation therapy; Rattus; Reproductive Medicine; Rewarming; Rhesus; Sales; Sampling; Scrotum; Services; Sex Functioning; Sickle Cell Anemia; Skin; Small Business Innovation Research Grant; Spermatogenesis; Stains; Structure; Sucrose; Surgeon; Surgical Replantation; Systemic Lupus Erythematosus; Systemic Scleroderma; Technology; Testicular Tissue; Testing; Testis; Testosterone; Therapeutic immunosuppression; Time; Tissue Banks; Tissue Viability; Tissues; Transplantation; Trehalose; base; boys; cancer diagnosis; cancer therapy; chemotherapy; childhood cancer survivor; commercialization; cytopenia; cytotoxic; cytotoxicity; formamide; implantation; improved; innovation; male; nanoparticle; nanowarming; novel; prepuberty; preservation; radio frequency; repaired; reproductive; reproductive organ; restoration; sperm cell; stem cells; vapor; ward; young adult

Thousands of young males under the age of 20 are diagnosed with cancer each year. The overall cure rates of these cancer patients are approaching 80%; therefore, the number of childhood cancer survivors is increasing over time. It is well documented that cancer treatments may result in temporary, long-term, or permanent gonadal failure in male cancer survivors. Certain benign hematological disorders, such as myelodysplasia, sickle cell disease, aplastic anemia, thalassemia major, and Fanconi anemia, and severe autoimmune diseases unresponsive to immunosuppressive therapy, such as juvenile idiopathic arthritis, juvenile systemic lupus erythematosus, systemic sclerosis and immune cytopenias, necessitate administration of high dose chemotherapy that may also impact testicles. In immature boys, in particular, storing testicular tissue prior to such cytotoxic treatments for future replantation could be an option. This proposal specifically addresses cryopreservation of pediatric testes. This male reproductive organ is contained within the scrotum, and is responsible for the production of sperm and the male hormone, testosterone that is pivotal for the maintenance of male sexual characteristics. As such, injury to the testes can render a young male infertile and needing administration of exogenous testosterone. Klinefelter’s syndrome patients would also benefit. Klinefelter’s syndrome the most common (1:500 new male born) genetic reason for male infertility, suffer from progressive germ cell loss and testis fibrosis with onset of puberty. We propose to remedy this by cryopreservation of testes that can be used for restoration of sexual function, fertility and hormonal balance in males before they are damaged. A single testis from each patient would be cryopreserved. This would permit either replantation of the patient’s intact testis, implantation of viable spermatogonial stem cells in the remaining testis, or ectopic implantation of functional spermatogonial stem cells. Each strategy would be capable of completing spermatogenesis and provide testosterone in a regulated way. We propose a novel innovative approach using ice-free vitrification to cryopreserve pediatric testes employing a young rat model. No animals will be sacrificed because we are using heterozygous and wild type culls from breeding colonies post-mortem. In this Phase I SBIR proposal in vitro methods and histology will be used to assess outcomes in three specific aims. Feasibility for progression to a Phase II SBIR grant proposal will be demonstrated by achievement of >80% cell viability by 3 methods and <20% apoptotic cells after 3 months of storage in vapor phase nitrogen. We have already assembled a world-class team of experts in surgery and reproductive medicine to guide the Company in Phase II and Phase III commercialization efforts.

每年有数千名20岁以下的年轻男性被诊断出患有癌症。这些癌症患者的总体治愈率接近80%;因此，儿童癌症幸存者的数量随着时间的推移而增加。据文献记载，癌症治疗可能导致男性癌症幸存者暂时，长期或永久性性腺衰竭。某些良性血液疾病，如骨髓增生异常、镰状细胞病、再生障碍性贫血、重型地中海贫血和范可尼贫血，以及对免疫抑制治疗无反应的严重自身免疫性疾病，如幼年特发性关节炎、幼年系统性红斑狼疮、系统性硬化症和免疫性血细胞减少症，需要给予高剂量化疗，这也可能影响睾丸。特别是在未成熟的男孩中，在这种细胞毒性治疗之前储存睾丸组织以备将来再植可能是一种选择。本提案专门针对儿科睾丸的冷冻保存。这个男性生殖器官包含在阴囊内，负责生产精子和男性激素，睾丸激素是维持男性性特征的关键。因此，睾丸损伤可导致年轻男性不育，需要给予外源性睾酮。Klinefelter综合征患者也将受益。Klinefelter综合征是最常见的（1：500新生男性）男性不育的遗传原因，患有进行性生殖细胞丢失和睾丸纤维化，伴随青春期的开始。我们建议通过冷冻保存睾丸来补救这一点，这些睾丸可用于在男性受损之前恢复性功能，生育力和激素平衡。每个病人的一个睾丸将被冷冻保存。这将允许患者的完整睾丸再植，在剩余的睾丸中植入活的精原干细胞，或异位植入功能性精原干细胞。每种策略都能够完成精子发生，并以可调节的方式提供睾酮。我们提出了一种新的创新方法，使用无冰玻璃化冷冻保存儿童睾丸采用年轻的大鼠模型。将不处死动物，因为我们使用的是来自死后繁殖群体的杂合和野生型剔除动物。在本I期SBIR提案中，将使用体外方法和组织学评估三个特定目标的结局。将通过3种方法实现>80%的细胞活力和在气相氮气中储存3个月后<20%的凋亡细胞来证明进展到II期SBIR资助提案的可行性。我们已经组建了一个世界级的外科和生殖医学专家团队，指导公司进行第二阶段和第三阶段的商业化工作。