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.

造血干细胞 (HSC) 移植 (HSCT) 被认为是治疗患有以下疾病的患者的潜在治疗选择 美国每年进行某些血液和骨髓癌症以及超过 20,000 例 HSCT。 然而，由于造血干细胞移植（HSCT）之前需要去除患病的骨髓，因此风险较高。 移植。直到移植的细胞植入并增殖到足以重建患者的免疫能力 系统中，患者被认为是中性粒细胞减少症，在此期间他们非常容易受到感染。此外 中性粒细胞减少症对患者可能存在危及生命的身体风险，因此需要较长的住院时间 在美国，造血干细胞移植的费用高达 40 万美元。目前的临床策略 试验的目的是移植更多的供体造血干细胞以缩短中性粒细胞减少期。然而，这些方法 全部依赖于供体细胞的离体培养，而供体细胞依赖于不完善的人造培养系统，可能会破坏 干细胞的正常功能，并且可能非常昂贵。忒修斯之船的创新解决方案是 一种在移植前短暂、一次性接触供体细胞的药物，可改善 体内扩张。我们的药物是核转录因子同源盒蛋白 B4 的专利突变体 (HOXB4(m))，促进 HSC 扩增而不分化。 HOXB4(m) 改善了降解 耐药性，这使我们能够克服 HOXB4 的几个缺点（例如蛋白质半衰期短和 与基因过度表达相关的骨髓增殖性疾病）。通过延长其细胞内半衰期， HOXB4(m) 具有实用性，可用于临床，同时可能避免由组成型引起的不良反应 过度表达。早期临床前数据表明用 HOXB4(m) 处理细胞可以扩展所有主要谱系 HSC 同时保持多能性。然而，目前 HOXB4(m) 的输送受到其穿透能力的限制。 细胞并定位于细胞核，即其生物活性的部位。 该 R61/R33 提案的目标是通过改进 HOXB4(m) 的核性能来提高 HOXB4(m) 的效率和安全性。 送货。这将通过执行 3 个目标来实现。在目标 1 (R61) 中，我们将执行一个库 筛选以确定 HOXB4(m) 细胞穿透肽和核靶向基序的最佳组合 核定位。在目标 2 (R33) 中，我们将评估最佳候选药物的体外功效，以选择先导药物 候选人。在目标 3 (R33) 中，我们将证明长期造血重建不会产生不良影响 或我们的主要候选者在小鼠 HSCT 研究中体内的 GVHD。成功完成R61/R33项目 将证明改进的 HOXB4(m) 候选药物在较低浓度下保持功效的可行性 和更短的暴露时间。该项目将为支持未来广泛的体内研究提供必要的数据 IND 提交所需的有效性和安全性研究。使用 HOXB4(m)，我们可以绕过原来的 克服 HOXB4 的障碍并改善体内扩增，通过缩短中性粒细胞减少期使 HSCT 患者受益 以减少感染、住院时间和费用。