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.

项目总结/摘要 免疫细胞疗法（ACT）是治疗癌症的一种有前途的治疗方法。然而，在这方面， ACT的最初成功仅限于嵌合抗原受体（CAR）-T细胞疗法， 恶性肿瘤。将这种细胞疗法应用于实体瘤受到缺乏可靶向肿瘤抗原的挑战， 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 可以克服这些挑战中的一些，因为它能够靶向细胞内蛋白质， 人白细胞抗原（HLA）复合物上的肽抗原。然而，大多数自然发生的 TCR对其肽-HLA靶标具有低亲和力。通过噬菌体展示或酵母展示工程化这些TCR 由于引入了不需要的交叉反应性和不良的结合， 亲和力和功能之间的关系该项目旨在解决ACT的每一个主要挑战，以便 有效地重新编程免疫系统以对抗实体瘤。 F99阶段的重点是TCR工程平台，用于创建、修改和分析 可以靶向肿瘤相关自身蛋白的TCR，毒性最小。在这种方法中，我首先提高T 细胞从天然库中识别一个相关的“外来”肽，该肽与天然库中的肽相差一个氨基酸。 自身肽然后，通过肽结合区的定向进化， 以将其特异性转向感兴趣的肿瘤自身抗原。我通过下面的例子证明了这种方法的价值： 病毒特异性TCR库的创建以及随后体外选择转换特异性的TCR 一个紧密相关的抗原决定基工程改造的TCR在配体识别后显示出稳健的T细胞活化， 具有与亲本受体相同或更高的效率。重要的是，工程改造的TCR没有显示出 与亲本TCR相比，额外的混杂性或脱靶特异性。剩下的目标是 我的论文项目是将这种方法应用于产生癌症反应性TCR。通过控制 由于TCR具有良好的特异性，这种方法将克服ACT的两个主要挑战，即增加 广泛的抗原可用于ACT，同时还最大限度地减少交叉反应性。 对于K 00阶段，我将把重点转移到解决抑制性肿瘤微环境， 通过开发新的合成受体并在小鼠模型中进行测试，我计划建造 在我的合成生物学背景下，实现新的高通量筛选，学习新的统计分析 方法，并获得使用体内小鼠癌症模型的经验。这些新方法加上 凭借我在遗传学、分子生物学和生物化学方面已经很强的背景， 靶向免疫治疗面临的最紧迫和最具挑战性的问题。在这个奖项的帮助下，我打算 继续我的研究贡献，成为癌症免疫治疗领域的领导者。