PLT+ Characterization and IND-Enabling Studies

PLT 表征和 IND 支持研究

• 批准号: 10223403
• 负责人: Bradford Dykstra
• 金额: $ 95.21万
• 依托单位: PLATELET BIOGENESIS, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10223403
• 关键词: Adhesions; Adult; Adverse reactions; Animal Model; Animals; Benchmarking; Biogenesis; Biological Assay; Biological Markers; Bioreactors; Birth; Blood; Blood Circulation; Blood Platelets; Blood Vessels; Bone Marrow; Cell Culture System; Cell Line; Cells; Clinical; Clinical Research; Clinical Trials; Coagulation Process; Consumption; Contracts; Dangerousness; Data; Dose; Emergency Situation; Equipment and supply inventories; Feedback; Freezing; Funding; Generations; Goals; Growth; HIV; Hand; Health; Hemorrhage; Hemostatic Agents; Human; Human Volunteers; Immunocompromised Host; In Vitro; Industry; Infusion procedures; Intravenous; Intravenous infusion procedures; Investigational Drugs; Investigational New Drug Application; Life; Malignant Neoplasms; Measures; Medical; Megakaryocytes; Metabolic; Microfluidics; Modeling; Monitor; Mus; Non obese; Operative Surgical Procedures; Phase; Phase I Clinical Trials; Physiological; Pilot Projects; Platelet Count measurement; Population; Positioning Attribute; Practice Guidelines; Pregnancy; Process; Production; Protocols documentation; Residual state; Rest; Risk; Safety; Sepsis; Serum; Small Business Innovation Research Grant; Source; Sterility; Survival Rate; Technology; Teratogens; Teratoma; Testing; Therapeutic; Thrombin; Time; Transcript; Transfusion; Transplantation; Transplantation Surgery; Undifferentiated; United States Food and Drug Administration; Virus Diseases; base; burn therapy; cancer therapy; cell bank; clinical practice; design; detection limit; diabetic; dosage; efficacy study; experimental study; first-in-human; good laboratory practice; in vivo; induced pluripotent stem cell; manufacturing process; meetings; nano-string; platelet function; platelet storage; pre-clinical; prevent; progenitor; repaired; research clinical testing; safety study; screening; stem cell biomarkers; therapeutic candidate; thrombogenesis; tumor; tumorigenesis; tumorigenic; validation studies; volunteer

PROJECT SUMMARY/ABSTRACT Platelet BioGenesis has developed a millifluidic bioreactor that reproduces key features of the adult bone marrow microenvironment to enable clinical-scale production of donor-independent platelets (PLT+) from human induced pluripotent stem cells (hiPSCs). Based on guidance from the U.S. Food & Drug Administration (FDA), the goal of this proposal is to perform safety and efficacy studies following good laboratory practice (GLP) principles to support a regulatory package that will permit clinical testing of our product. Platelets are essential blood components responsible for clot formation and blood vessel repair. Low platelet count is a dangerous consequence of cancer treatment, transplant, and surgery, and platelets are a critical first-line therapy to prevent uncontrolled bleeding. Transfusion units are derived exclusively from human volunteer donors and must be stored at ≥22°C to avoid irreversible activation/aggregation. Risk of bacterial growth limits shelf life to 5 days, 3 of which are consumed by screening and transport. Because of these limitations, blood centers often only have a 1.5-day inventory that is quickly depleted by emergencies [1,2]. We have developed a 2-step ex vivo production platform in which hiPSCs (replenishable progenitors that can be frozen for years) undergo differentiation into pre- megakaryocytes (preMK+) that are banked and used for on-demand PLT+ generation in our bioreactor. In our Phase II project, we successfully generated and characterized functional preMKs+ and PLTs+ from a qualified hiPSC line [3,4] using an established serum/feeder-free, scalable current good manufacturing practice (cGMP)-compliant process [5]. After confirming PLT+ quality and function, we verified hemostatic and thrombogenic potentials in vivo. Building on these results, this Phase IIB SBIR proposal outlines three specific aims to perform GLP studies to 1) establish PLT+ release criteria and storage profile, 2) assess teratoma risk in vivo, and 3) measure circulation and clearance time in vivo. For all aims, human donor platelets will serve as a physiological benchmark for comparison. Completion of these aims will facilitate submission of an Investigational New Drug (IND) application. Aim 1. Establish PLT+ release quality and storage profile. We will evaluate PLT+ biomarkers, function, metabolic activity, and sterility under FDA-approved storage conditions. Aim 2. Assess teratoma risk in mice. Following GLP guidelines, we will determine whether any risk of tumorigenic growth is associated with PLT+ product in immunocompromised (NSG) mice. Aim 3. Assess PLT+ circulation and clearance times in mice. We will assess the circulation and clearance of PLT+ in immunocompromised (NSG) mice.

项目总结/摘要 Platelet BioGenesis开发了一种毫流体生物反应器，可以再现成年人的关键特征。 能够临床规模生产非供体依赖性血小板的骨髓微环境 (PLT+）来自人诱导多能干细胞（hiPSC）。根据美国食品安全局的指导， 与美国食品药品监督管理局（FDA），该提案的目标是进行安全性和有效性研究 遵循药物非临床研究质量管理规范（GLP）原则，以支持允许 我们产品的临床试验。血小板是血液中的重要成分， 形成和血管修复。低血小板计数是癌症的危险后果 治疗、移植和手术，血小板是预防糖尿病的关键一线疗法。 无法控制的出血输血单位仅来自人类志愿者供体， 必须在≥ 22 ° C下储存，以避免不可逆的活化/聚集。细菌生长极限的风险 保质期为5天，其中3天用于筛选和运输。因为这些 由于血液中心的局限性，血液中心通常只有1.5天的库存， 紧急情况[1，2]。我们已经开发了两步离体生产平台，其中hiPSC （可以冷冻多年的可腐烂的祖细胞）经历分化成前 巨核细胞（preMK+）被储存并用于按需生成PLT+， 生物反应在我们的第二阶段项目中，我们成功地生成并表征了功能性preMK + 和PLT+来自合格的hiPSC系[3，4]，使用已建立的无血清/饲养层、可扩展电流 良好生产规范（cGMP）合规过程[5]。在确认PLT+质量和功能后， 我们在体内验证了止血和血栓形成的潜力。在这些结果的基础上，本阶段IIB SBIR提案概述了进行GLP研究的三个具体目标，以1）建立PLT+放行 标准和储存概况，2）评估体内畸胎瘤风险，和3）测量循环和 体内清除时间。对于所有的目标，人类捐献的血小板将作为一个生理基准 以供比较。完成这些目标将有助于提交新的研究报告。 药物（IND）申请。 目标1.建立PLT+放行质量和储存概况。我们将评估PLT+生物标志物， 功能、代谢活性和无菌性。 目标2.评估小鼠畸胎瘤风险。根据GLP指南，我们将确定是否存在任何风险 在免疫受损（NSG）小鼠中，致瘤性生长与PLT+产物相关。 目标3.评估小鼠中的PLT+循环和清除时间。我们将评估发行量 和免疫受损（NSG）小鼠中PLT+的清除。