Improving the activity of CAR T cells for acute myeloid leukemia

提高CAR T细胞治疗急性髓系白血病的活性

• 批准号: 10740585
• 负责人: Miriam Y Kim
• 金额: $ 24.9万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10740585
• 关键词: Acute Myelocytic Leukemia; Allogenic; Antigen Targeting; Attention; B-Lymphocytes; Bioinformatics; Biology; Biometry; Bispecific Antibodies; C57BL/6 Mouse; CAR T cell therapy; CD19 gene; CRISPR/Cas technology; Cell Count; Cell Therapy; Cell physiology; Cells; Clinical; Complex; Data; Dendritic Cells; Development Plans; Disease; Disease Outcome; Effector Cell; Engineering; Environment; Environmental Risk Factor; Exposure to; Flow Cytometry; Foundations; Future; Gene Transfer; Goals; Harvest; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic System; Hematopoietic stem cells; Human; Immune; Immune system; Immunocompetent; Immunology; Immunotherapy; Impairment; In Vitro; Individual; Infection; Investigation; Learning; Leukemic Cell; Macrophage; Mediator; Mentors; Methods; Modems; Mus; Myelogenous; Myeloid Cells; Myeloproliferative disease; Nature; Patients; Phenotype; Population; Research; Research Personnel; Resistance; T-Lymphocyte; TCR Activation; Time; Tissues; Toxic effect; Training; Universities; Viral Genes; Washington; Work; acute myeloid leukemia cell; antigen-specific T cells; antitumor effect; cancer immunotherapy; career; career development; cell behavior; cellular engineering; chimeric antigen receptor; chimeric antigen receptor T cells; disorder control; improved; improved outcome; in vivo; innovation; knockout gene; leukemia; mouse model; negative affect; novel; research and development; response; single-cell RNA sequencing; skills; stem cell genes; success; targeted treatment; therapy outcome; tumor

Project Summary Acute myeloid leukemia (AML) has thus far proven resistant to T cell redirecting therapies such as bispecific antibodies or chimeric antigen receptor (CAR) T cells, which is unexpected given the proven success of these in other hematologic malignancies. Preliminary studies show that the anti-tumor activity of CD19-targeting CAR T cells (CART19) is enhanced in the presence of myeloid cells, and that targeting these myeloid cells with CD33-targeting CART cells (CART33) leads to diminished long-term anti-tumor effect. Furthermore, removing CD33 from normal myeloid cells improves CART33 expansion and disease control. Based on these findings, the central hypothesis of this proposal is that bystander myeloid cells enhance CART cell activity, and directly targeting myeloid cells compromises this effect. The objective of this proposal is to delineate how myeloid cells influence CAR T cell behavior when they are targets of therapy, as compared to when they are simply bystanders, so that we can intervene within these interactions and improve CAR T cell therapy for AML. This will be achieved through the following specific aims: 1) Determine how bystander myeloid cells modulate CAR T cell function; 2) Determine how targeting myeloid cells changes CAR T cell profile. To this end, hematopoietic stem cells (HSCs) will be engineered through either viral gene transfer or CRISPR/Cas9 gene knockout to generate myeloid cells that are positive or negative for the antigen targeted by the CAR. The effects of wild-type or genetically modified myeloid cells on CAR T cells will be interrogated by in vitro culture and in vivo mouse models, using flow cytometry, single-cell RNA-seq, and functional studies. The innovations of this project are that it draws attention to host environmental factors that influence CAR T cell activity, and advances the concept that the nature of the cell being targeted can influence CAR T cell behavior. This research is significant because it will contribute to a better understanding of how CAR T cells work with the immune environment, and illuminate methods to intervene within these interactions to improve the outcomes of therapy. The long-term goal of this proposal is to establish the applicant Dr. Miriam Kim's career as an independent researcher focused on developing novel cell therapies for AML. The proposed research and career development plan will provide Dr. Kim with training in HSC/myeloid biology, immunology, and bioinformatics. Her primary mentor, Dr. John DiPersio, and co-mentors Drs. Robert Schreiber, Todd Fehniger and Carl DeSelm, offer complementary expertise in cell engineering and immunotherapy. Additionally, Dr. Kim's collaborators Dr. Li Ding and Dr. Feng Gao will contribute to developing her skills in bioinforrnatics and biostatistics. Furthermore, Washington University provides an ideal environment for Dr. Kim to successfully establish herself as an independent investigator.

项目摘要 急性髓性白血病（AML）迄今已被证明对T细胞重定向疗法如双特异性免疫疗法具有抗性。 抗体或嵌合抗原受体（CAR）T细胞，这是出乎意料的，因为这些方法已经证明是成功的。 其他血液恶性肿瘤。初步研究表明，靶向CD 19的CAR的抗肿瘤活性 T细胞（CART 19）在骨髓细胞的存在下增强，并且靶向这些骨髓细胞的CART 19在骨髓细胞的存在下增强。 靶向CD 33的CART细胞（CART 33）导致长期抗肿瘤作用减弱。此外，去除 来自正常骨髓细胞的CD 33改善CART 33扩增和疾病控制。根据这些发现， 该建议的中心假设是旁观者骨髓细胞增强CART细胞活性，并直接 靶向骨髓细胞损害了这种效果。这项提议的目的是描述骨髓细胞如何 当它们是治疗的靶点时，与当它们仅仅是 因此，我们可以干预这些相互作用，并改善AML的CAR T细胞治疗。这 将通过以下具体目标实现：1）确定旁观者骨髓细胞如何调节CAR T细胞功能; 2）确定靶向骨髓细胞如何改变CAR T细胞谱。为此目的， 造血干细胞（HSC）将通过病毒基因转移或CRISPR/Cas9基因工程化， 敲除以产生对CAR靶向的抗原呈阳性或阴性的骨髓细胞。的 野生型或遗传修饰的骨髓细胞对CAR T细胞的影响将通过体外培养来研究 和体内小鼠模型，使用流式细胞术，单细胞RNA-seq和功能研究。的创新 该项目的一个重要特点是，它引起了人们对影响CAR T细胞活性的宿主环境因素的关注， 提出了这样的概念，即靶向细胞的性质可以影响CAR T细胞的行为。这 这项研究意义重大，因为它将有助于更好地了解CAR T细胞如何与细胞一起工作。 免疫环境，并阐明干预这些相互作用的方法，以改善免疫系统的结果。 疗法本提案的长期目标是将申请人Miriam Kim博士的职业生涯确立为 独立研究人员专注于开发AML的新型细胞疗法。拟议的研究和 职业发展计划将为金博士提供HSC/骨髓生物学，免疫学和 生物信息学她的主要导师，博士约翰DiPersio，和共同导师博士罗伯特Schreiber，托德Fehniger 和Carl DeSelm，提供细胞工程和免疫治疗方面的互补专业知识。此外，博士。 Kim的合作者Li Ding博士和Feng Gao博士将致力于发展她在生物信息学方面的技能， 生物统计学此外，华盛顿大学为金博士成功地 让自己成为一名独立调查员