Investigating the Spatial Dynamics of Intratumoral Immune Cells in Breast Cancer

项目摘要 虽然免疫疗法使某些恶性肿瘤的治疗发生了革命性的变化，但它尚未产生同样的效果 会导致乳腺癌。这是惊人的，因为有大量的肿瘤内免疫细胞 会影响疾病的结果。有证据表明，这些白细胞处于战略地位，可以执行他们的 然而，对它们在肿瘤进展和消退过程中的空间动力学知之甚少。我的目标是 使用最新的进展详细描述这些时空动力学的全面和机械图景 高维原位成像。我假设随着肿瘤的发展，免疫抑制细胞会阻止 促进抗肿瘤免疫的免疫细胞组织结构和细胞间通讯 当肿瘤消退时，这种表型会逆转。这将通过描述空间特征来进行评估 主要白细胞的组织、细胞间相互作用、激活/抑制标志物和细胞因子的产生 病程中的原位血统。我的初步数据表明，我可以识别出这些主要的大部分 乳腺肿瘤内的免疫细胞类型，包括巨噬细胞、树突状细胞和T细胞的亚群。 此外，我发现巨噬细胞的两个亚群有不同的定位：一个是肿瘤内的。 另一种几乎完全发生在瘤周。这种瘤周巨噬细胞群密切地相互作用 T调节细胞，这对乳腺癌的免疫抑制很重要。因此，我假设 瘤周FDMs通过TCR-MHC II空间组织与T调节细胞相互作用，从而激活 Tregs和偏向免疫抑制微环境的肿瘤。为了测试这一点，我将使用Genetic 以肿瘤分期特异的方式耗尽这些群体的小鼠模型，并评估其对肿瘤的影响 生长、存活和对临床治疗的反应。总体而言，这项建议旨在了解 乳腺癌中白细胞的动态变化，这将揭示关键的细胞-细胞相互作用，组织结构， 以及需要被破坏/促进的细胞通信网络，以重新参与抗肿瘤 免疫，促进有效的免疫疗法的发展。

PROJECT ABSTRACT While immunotherapy has revolutionized the treatment of certain malignancies it has yet to yield the same results in breast cancer. This is striking, given that there is an abundance of intra-tumoral immune cells that affect disease outcomes. Evidence suggests that these leukocytes are strategically positioned to execute their functions and yet little is known about their spatial dynamics over tumor progression and regression. I aim to detail a comprehensive and mechanistic picture of these spatiotemporal dynamics using recent advances in high-dimensional in situ imaging. I hypothesize that as tumors progress, immunosuppressive cells will block immune cell organizational structures and cell-cell communications that promote antitumor immunity and that this phenotype will be reversed upon tumor regression. This will be assessed by characterizing the spatial organization, cell-cell interactions, activation/inhibition markers, and cytokine production of the main leukocyte lineages in situ over disease course. My preliminary data demonstrate that I can identify most of these major immune cell types at once within breast tumors, including subsets of macrophages, dendritic cells, and T cells. In addition, I found that two subsets of macrophages have differential localization: one which is intra-tumoral and another that is almost exclusively peritumoral. This peritumoral macrophage population closely interacts with T regulatory cells, which are important for immunosuppression in breast cancer. As such, I hypothesize that peritumoral FDMs spatially organize to interact with T regulatory cells via TCR-MHC II, thereby activating Tregs and skewing tumors toward an immunosuppressed microenvironment. To test this, I will use genetic mouse models to deplete these populations in a tumor stage-specific manner and assess the impact on tumor growth, survival, and response to clinical therapies. Overall, this proposal aims to understand the spatial dynamics of leukocytes in breast cancer, which will reveal key cell-cell interactions, organizational structures, and cellular communication networks that need to be disrupted/promoted in order to reengage antitumor immunity, facilitating the development of effective immunotherapies.