Preclinical development of a nuclear-targeting biologic that safely increases stem cell expansion in vivo to accelerate recovery from neutropenia after chemotherapy and bone marrow transplant

临床前开发一种核靶向生物制剂，可安全地增加干细胞体内扩增，以加速化疗和骨髓移植后中性粒细胞减少症的恢复

• 批准号: 10685290
• 负责人: Jeremy A Elser
• 金额: $ 23.03万
• 依托单位: SHIP OF THESEUS, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-17 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685290
• 关键词: ANXA5 gene; Acceleration; Adverse effects; Adverse event; Aftercare; Apoptosis; Apoptotic; Biological; Biological Assay; Blood; Bone Marrow; Bone Marrow Diseases; Bone Marrow Transplantation; CD34 gene; Cell Nucleus; Cell Transplantation; Cells; Chemotherapy and/or radiation; Chimeric Proteins; Chimerism; Clinic; Clinical; Clinical Trials; Complex; Consumption; Data; Disease; Dose; Dyes; Engraftment; Excision; Exploratory/Developmental Grants Phase II; Exposure to; Female; Fluorescence Microscopy; Future; Gel; Genetic; Goals; Half-Life; Hela Cells; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Homeodomain Proteins; Hospitalization; Hour; Human; Immune system; Immunodeficient Mouse; In Vitro; Infection; Inflammatory; Inpatients; Intravenous infusion procedures; Lead; Legal patent; Libraries; Life; Malignant Bone Marrow Neoplasm; Measures; Methodology; Methods; Modeling; Monitor; Mucositis; Mus; Myelogenous; Myeloproliferative disease; Necrosis; Neutropenia; Nuclear; Patient-Focused Outcomes; Patients; Penetration; Peptides; Pharmaceutical Preparations; Phase; Predisposition; Proliferating; Proteins; Recovery; Resistance; Risk; Safety; Scanning; Site; Source; Stains; System; Therapeutic; Time; Toxic effect; Toxicology; Transplant Recipients; Transplantation; Transplantation Conditioning; candidate identification; candidate selection; chemotherapy; clinical investigation; comparison control; conditioning; cost; cytotoxicity; efficacy evaluation; efficacy study; experience; hematopoietic stem cell expansion; high risk; improved; in vivo; infection risk; innovation; lead candidate; male; man; manufacture; mutant; novel; overexpression; peripheral blood; post-transplant; pre-clinical; preclinical development; reconstitution; safety study; screening; stem cell expansion; stem cell function; transcription factor; uptake

Hematopoietic stem cell (HSC) transplants (HSCTs) are considered a potentially curative option for patients with certain cancers of the blood and bone marrow and over 20,000 HSCTs are performed in the US each year. However, HSCTs are associated with high risk due to the required removal of diseased bone marrow before transplantation. Until the transplanted cells engraft and proliferate enough to reconstitute the patient’s immune system, the patient is considered neutropenic during which they are highly susceptible to infection. In addition to the potentially life-threatening physical risk to the patient, neutropenia necessitates long hospitalization time that can in part drives the expensive cost of HSCTs ranging up to $400,000 in the US. A current strategy in clinical trials is to transplant larger pools of donor HSCs to shorten the neutropenic phase. However, these approaches all rely on ex vivo culturing of donor cells that depend on imperfect man-made culture systems that may disrupt the proper function of the stem cells and can be prohibitively expensive. Ship of Theseus’ innovative solution is a drug that will be used for a brief, one-time exposure to donor cells prior to transplantation that improves expansion in vivo. Our drug is a patented mutant of nuclear transcription factor, Homeobox protein B4 (HOXB4(m)), which promotes HSC expansion without differentiation. HOXB4(m) has improved degradation resistance, which enables us to overcome several shortcomings of HOXB4 (e.g., short protein half-life and myeloproliferative disorders associated with genetic overexpression). By elongating its intracellular half-life, HOXB4(m) is practical for clinical use while potentially avoiding adverse effects induced by constitutive overexpression. Early preclinical data indicating treating cells with HOXB4(m) can expand all major lineages of HSCs while maintaining multipotency. However, current delivery of HOXB4(m) is limited by its ability to penetrate cells and localize in the nucleus, the site of its bioactivity. The goal of this R61/R33 proposal is to improve efficiency and safety of HOXB4(m) by improving its nuclear delivery. This will be accomplished through the execution of 3 aims. In Aim 1 (R61), we will perform a library screen to identify optimal combinations of cell penetrating peptide and nuclear targeting motifs for HOXB4(m) nuclear localization. In Aim 2 (R33), we will assess in vitro efficacy of the top candidates to select a lead candidate. In Aim 3 (R33), we will demonstrate long-term hematopoietic reconstitution without adverse effects or GVHD of our lead candidate in vivo in a mouse HSCT study. Successful completion of this R61/R33 program will demonstrate feasibility of an improved HOXB4(m) candidate that maintains efficacy at lower concentrations and briefer exposure periods. This project will provide the necessary data to support future extensive in vivo efficacy and safety studies necessary for an IND submission. With HOXB4(m), we can circumvent the original hurdles of HOXB4 and improve expansion in vivo to benefit HSCT patients by shortening the neutropenic phase to reduce infection, hospitalization time, and cost.

对于患者而言 每年在美国进行某些血液和骨髓癌和超过20,000个HSCT。 但是，由于需要去除患病的骨髓，因此HSCTS与高风险有关 移植。直到移植的细胞植入并扩散到足以重建患者的免疫力 系统，患者被认为是中性的，在此期间，他们非常容易感染。此外 可能对患者的潜在威胁生命的身体风险，中性粒细胞减少必要的长期住院时间 可以部分地驱动美国HSCT的昂贵成本，在美国最高40万美元。当前的临床策略 试验是将较大的供体HSC池移植以缩短中性相。但是，这些方法 所有这些都依赖于依赖不完美的人造培养系统的供体细胞的离体培养 干细胞的适当功能，可以禁止昂贵。 Theseus的创新解决方案是 在移植之前，将用于短暂，一次性接触供体细胞的药物，以改善 体内扩展。我们的药物是核转录因子的专利突变体，同型蛋白B4 （HOXB4（M）），它促进HSC扩展而无需分化。 HOXB4（M）改善了降解 阻力，这使我们能够克服hoxb4的几个缺点（例如，蛋白质半衰期和 与遗传过表达相关的骨髓增生性疾病）。通过延长细胞内半衰期， HOXB4（M）在临床上是实用的，同时有可能避免一致性引起的不良反应 过表达。早期的临床前数据表明用HOXB4（M）处理细胞可以扩展所有主要谱系 HSC在维持多稳定性的同时。但是，Hoxb4（M）的当前输送受到其穿透能力的限制 细胞并定位在细胞核中，其生物活性部位。 该R61/R33提案的目的是通过改善其核能提高HOXB4（M）的效率和安全性 送货。这将通过执行3个目标来实现。在AIM 1（R61）中，我们将执行一个库 屏幕以识别HOXB4（M）的细胞穿透性肽和核靶标基序的最佳组合 核定位。在AIM 2（R33）中，我们将评估顶级候选者的体外效率以选择铅 候选人。在AIM 3（R33）中，我们将展示长期造血重建而不会产生不利影响 在小鼠HSCT研究中，我们的铅候选者的GVHD。成功完成此R61/R33程序 将证明提高的HOXB4（M）候选者的可行性，该候选者以较低的浓度保持效率 和英国曝光期。该项目将提供必要的数据，以支持未来的大量体内 提交IND所需的功效和安全性研究。使用Hoxb4（m），我们可以绕过原始 HOXB4的障碍并改善体内扩张，以使HSCT患者受益于中性期。 减少感染，住院时间和成本。