Cellular engineering to improve the efficacy and specificity of targeted immunotherapy

细胞工程提高靶向免疫疗法的功效和特异性

• 批准号: 10304452
• 负责人: Nouran Saied Abdelfattah
• 金额: $ 3.54万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10304452
• 关键词: Address; Adoptive Cell Transfers; Adverse event; Affect; Affinity; Amino Acids; Animal Model; Antigen Targeting; Antigens; Autoantigens; Award; B-Lymphocytes; Binding; Biochemistry; Bypass; CAR T cell therapy; CD8-Positive T-Lymphocytes; Cancer Model; Cancer cell line; Cell Therapy; Cessation of life; Clinical Trials; Collection; Complex; Coupled; Cytomegalovirus; Development; Directed Molecular Evolution; Engineering; Ensure; Epitopes; FDA approved; Face; Generations; Genetic; Genetic Engineering; Goals; HLA Antigens; Hematologic Neoplasms; Hematopoietic Neoplasms; Human Genome; Immune signaling; Immune system; Immunotherapy; In Vitro; Individual; Infiltration; Lead; Learning; Libraries; Ligands; Logic; Malignant Neoplasms; Methods; Modification; Molecular Biology; Peptides; Phage Display; Phase; Population; Protein Engineering; Proteins; Research; Research Project Grants; Research Proposals; Residual state; Signal Transduction; Solid Neoplasm; Specificity; Statistical Data Interpretation; Surface Antigens; T-Cell Activation; T-Cell Antigen Receptor Specificity; T-Cell Immunologic Specificity; T-Cell Proliferation; T-Cell Receptor; T-Cell Receptor Genes; T-Lymphocyte; Testing; Therapeutic; Tissues; Toxic effect; Training; Tumor Antigens; Tumor-infiltrating immune cells; Viral; Work; Yeasts; autoreactive T cell; base; blood treatment; cancer immunotherapy; cancer testis antigen; cancer therapy; cell killing; cellular engineering; chimeric antigen receptor; chimeric antigen receptor T cells; clinical efficacy; combat; cross reactivity; design; engineered T cells; exhaustion; experience; gene therapy; high throughput screening; improved; in vivo; in vivo Model; insight; interest; mouse model; neoplastic cell; novel; novel strategies; novel therapeutics; receptor; response; screening; success; synthetic biology; systemic toxicity; tool; tumor; tumor microenvironment; tumor specificity; tumor-immune system interactions; virome

Project Summary/Abstract Adoptive cell therapy (ACT) is a promising therapeutic approach for the treatment of cancer. However, the initial success of ACT has been limited to chimeric antigen receptor (CAR)-T cell therapies for hematological malignancies. Applying this cell therapy to solid tumors is challenged by the lack of targetable tumor antigens, the severe systemic toxicity and the suppressive tumor microenvironment. T cell receptor (TCR) gene therapy can overcome some of these challenges because it enables targeting of intracellular proteins presented as peptide antigens on the human leukocyte antigen (HLA) complex. However, the majority of naturally occurring TCRs are of low-affinity to their peptide-HLA targets. Engineering these TCRs via phage display or yeast display for higher affinity is complicated by the introduction of unwanted cross-reactivity and the poor association between affinity and function. This project seeks to tackle each of the major challenges of ACT in order to effectively reprogram the immune system to combat solid tumors. The F99 phase is focused on a TCR engineering platform for the creation, modification, and profiling of TCRs that can target tumor-associated self-proteins with minimal toxicity profiles. In this approach, I first raise T cells from the natural repertoire that recognize a related ‘foreign’ peptide that differs by one amino acid from the self-peptide. Then, I modulate the fine specificity of the TCR by directed evolution of the peptide binding region to switch its specificity towards the tumor self-antigen of interest. I demonstrate the value of this approach by the creation of libraries of viral-specific TCRs and the subsequent in vitro selection of TCRs that switched specificity to a closely related epitope. The engineered TCRs showed robust T-cell activation after ligand recognition and are of equal or higher efficiency than the parental receptor. Importantly, the engineered TCRs displayed no additional promiscuity or off-target specificities as compared to the parental TCRs. The goal for the remainder of my dissertation project is to apply this approach to the generation of cancer reactive TCRs. By controlling the fine specificity of TCRs, this approach will overcome two of the major challenges of ACT, namely increasing the breadth of antigens that can be used for ACT while also minimizing cross-reactivities. For the K00 phase, I will shift my focus to addressing the suppressive tumor microenvironment that surrounds solid tumors by developing novel synthetic receptors and testing them in mouse models. I plan to build upon my synthetic biology background to implement novel high-throughput screens, learn new statistical analysis methods, and gain experience working with in vivo mouse models of cancer. These new approaches, coupled with my already strong background in genetics, molecular biology and biochemistry will allow me to address the most pressing and challenging issues facing targeted immunotherapies. With the aid of this award, I intend to continue my research contribution to become a leader in the field of cancer immunotherapy.

项目摘要/摘要 收养细胞疗法（ACT）是癌症治疗的有前途的治疗方法。然而， ACT的最初成功仅限于血液学的嵌合抗原受体（CAR）-T细胞疗法 恶性肿瘤。缺乏可靶向的肿瘤抗原，将这种细胞疗法应用于实体瘤的挑战 严重的全身毒性和抑制性肿瘤微环境。 T细胞受体（TCR）基因治疗 可以克服其中一些挑战，因为它可以针对呈现的细胞内蛋白 人白细胞抗原（HLA）复合物上的肽抗原。但是，大多数自然发生 TCR对其肽-HLA靶标的亲和力低。通过噬菌体展示或酵母显示器来工程这些TCR 对于更高的亲和力而言，由于引入不良的交叉反应性和较差的关联而变得复杂 在亲和力和功能之间。该项目旨在应对行为的每一个主要挑战 有效地重新编程免疫系统以对抗实体瘤。 F99阶段专注于TCR工程平台，用于创建，修改和分析 可以靶向具有最小毒性特征的肿瘤相关的自蛋白的TCR。在这种方法中，我首先提出 来自自然曲目的细胞识别相关的“异物”肽，该肽与一个氨基酸不同 自肽。然后，我通过肽结合区域的定向演化来调节TCR的精细特异性 将其特异性转向感兴趣的肿瘤自我抗原。我证明了这种方法的价值 创建病毒特异性TCR的库以及随后的体外选择TCR，以切换特异性 与密切相关的epoisodeTope。工程的TCR在配体识别后显示出强大的T细胞激活和 比父母接收者具有相等或更高的效率。重要的是，工程的TCR不显示 与父母TCR相比，其他滥交或脱离目标规格。其余的目标 我的论文项目是将这种方法应用于癌症反应性TCR的产生。通过控制 TCR的良好特异性，这种方法将克服两个行为的主要挑战，即增加 可用于行动的抗原的广度，同时还可以最大程度地减少交叉反应性。 对于K00阶段，我将重点转移到解决抑制性肿瘤微环境中 通过开发新型的合成受体并在小鼠模型中测试它们来包围实体瘤。我打算建造 在我的合成生物学背景下，以实现新颖的高通量屏幕，学习新的统计分析 方法，并获得与体内小鼠癌症模型一起工作的经验。这些新方法，耦合 凭借我已经很强的遗传学背景，分子生物学和生物化学将使我能够解决 最紧迫和挑战针对性免疫疗法面临的问题。在这个奖项的帮助下，我打算 继续我的研究贡献成为癌症免疫疗法领域的领导者。