Three-dimensional organoid models to study breast cancer progression

研究乳腺癌进展的三维类器官模型

基本信息

批准号：
10581806
负责人：
Shilpa Sant
金额：
$ 42.3万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10581806
关键词：
3-Dimensional AKT Signaling Pathway Address Agreement Antitumor Response Award Biomedical Engineering Breast Cancer Detection Breast Cancer Patient CCL2 gene CXCL10 gene CXCR3 gene Cancer cell line Carcinoma in Situ Cause of Death Cell Line Cell secretion Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Coupled Development Diagnosis Drug Screening Duct (organ) structure E-Cadherin Engineering Environment Epidermal Growth Factor Receptor Exhibits Experimental Models Fibronectins Genetic Heterogeneity Hybrids Hypoxia IL7 gene IL8 gene Image Analysis In Vitro Individual Interleukin-10 Interleukin-15 Interleukin-6 Invaded Knock-in Knowledge Label Lead Left Link Maintenance Malignant - descriptor Malignant Neoplasms Mammography Matrix Metalloproteinases Metabolic stress Metastatic breast cancer Micro Array Data Microarray Analysis Modeling Neoplasm Metastasis Noninfiltrating Intraductal Carcinoma Organoids Outcome PI3K/AKT PIK3CG gene Parents Pathway interactions Patients Peripheral Phenotype Pleural Pleural effusion disorder Prognostic Marker Proteins Reproducibility Resolution Role Sampling Signal Pathway Signal Transduction Site Sorting Spatial Distribution Stimulus Stress Structure System Testing Therapeutic Time Variant Vimentin Visualization Woman Work breast cancer progression clinically relevant confocal imaging cytokine deep learning deep learning algorithm design effective therapy gene regulatory network genetic signature genomic profiles imaging approach improved in vitro Model in vivo infiltrating duct carcinoma innovation malignant breast neoplasm meter migration neoplastic cell new therapeutic target novel overtreatment paracrine premalignant prevent progression risk rho GTP-Binding Proteins risk prediction therapy development treatment strategy tumor tumor heterogeneity

项目摘要

Abstract Approximately 20% of breast cancers detected through mammography are pre-invasive Ductal Carcinoma in situ (DCIS). If left untreated, approximately 20-50% of DCIS will progress to more deadly Invasive Ductal Carcinoma (IDC). No prognostic biomarkers can reliably predict the risk of progression from DCIS to IDC. Similar genomic profiles of matched pre-invasive DCIS and IDC suggests that the progression is not driven by genetic aberrations in DCIS cells, but microenvironmental factors, such as hypoxia and metabolic stress prevalent in DCIS, may drive the transition. We need innovative models to investigate how to halt steps of DCIS progression to invasive phenotypes and subsequent metastasis from the primary site. This proposal directly addresses this unmet need by developing a novel three-dimensional in vitro organoid model that recapitulates key hallmarks of DCIS to IDC progression: tumor-size induced hypoxia and metabolic stress, tumor heterogeneity and spontaneous emergence of migratory phenotype in the same parent cells without any additional stimulus. A tangible advantage of the proposed organoid models is the ability to precisely and reproducibly study how the hypoxic microenvironment induces tumor migration in real time and in isolation from non-tumor cells present in vivo, providing unique opportunity to define tumor-intrinsic mechanisms of DCIS to IDC progression. During July 2018-Feb 2022 ESI MERIT Award period, we have shown that inhibition of tumor-secreted factors effectively halts organoid migration, while inhibition of hypoxia is effective only within a time window and is compromised by tumor-to-tumor variation, supporting our notion that hypoxia initiates migratory phenotypes but does not sustain it. We have also analyzed secretome from metastatic breast cancer pleural effusion showing significantly higher levels of CCL2/MCP1, CXCL10/IP10, IL-6, IL-8, regulatory IL-10, and IL-7 and IL-15. Strategies to neutralize these key cytokines may generate anti-tumor responses in the pleural environment. Microarray analysis of hypoxia-induced migration and secretome-induced migration suggested role of Rho GTPase and PI3K/AKT signaling pathways in maintaining migration. Our results show that hypoxic organoid models exhibit partial EMT signatures as early as day 1, which is maintained as these non-migratory organoids transition to migratory phenotypes. During the two-year extension period, we will continue 1) to optimize our DCIS models incorporating ductal structure and other components from DCIS microenvironments; 2) to test new mechanisms linking tumor-intrinsic hypoxia, partial/hybrid EMT and collective migration; 3) to inhibit signaling mechanisms to halt emergence of migratory phenotypes. The successful completion of the proposed work will provide answers to two fundamental questions in the progression of invasive breast cancer: 1) What causes some DCIS cells to become migratory and develop into invasive tumors? 2) How and where does the migratory phenotype (IDC) emerge? The mechanistic understanding gained from these studies will improve diagnosis, lead to the development of treatment strategies to arrest invasion at the pre-malignant stage, and thus prevent patient overtreatment.

抽象的在通过乳房 X 光检查检测到的乳腺癌中，大约 20% 是浸润前导管癌原位（DCIS）。如果不及时治疗，大约 20-50% 的 DCIS 将发展为更致命的侵入性导管癌症（IDC）。没有预后生物标志物可以可靠地预测从 DCIS 进展为 IDC 的风险。相似的匹配的侵入前 DCIS 和 IDC 的基因组图谱表明，进展并非由遗传驱动 DCIS 细胞中存在畸变，但微环境因素（例如缺氧和代谢应激）在 DCIS 细胞中普遍存在 DCIS 可能会推动这一转变。我们需要创新模型来研究如何阻止 DCIS 进展侵袭性表型和随后从原发部位的转移。该提案直接针对通过开发一种新颖的三维体外类器官模型来满足这一未满足的需求，该模型概括了关键 DCIS 向 IDC 进展的标志：肿瘤大小引起的缺氧和代谢应激、肿瘤异质性以及在没有任何额外刺激的情况下在相同亲本细胞中自发出现迁移表型。一个所提出的类器官模型的明显优势是能够精确且可重复地研究缺氧微环境实时诱导肿瘤迁移，并且与存在于肿瘤细胞中的非肿瘤细胞隔离。体内，为定义 DCIS 到 IDC 进展的肿瘤内在机制提供了独特的机会。在 2018 年 7 月至 2022 年 2 月 ESI 优异奖期间，我们发现抑制肿瘤分泌因子有效地阻止类器官迁移，而抑制缺氧仅在一个时间窗口内有效，并且受到肿瘤间变异的影响，支持我们的观点，即缺氧启动迁移表型，但不支持它。我们还分析了转移性乳腺癌胸腔积液的分泌组，显示 CCL2/MCP1、CXCL10/IP10、IL-6、IL-8、调节性 IL-10、IL-7 和 IL-15 水平显着升高。中和这些关键细胞因子的策略可能会在胸膜环境中产生抗肿瘤反应。缺氧诱导的迁移和分泌蛋白组诱导的迁移的微阵列分析表明 Rho 的作用 GTPase 和 PI3K/AKT 信号通路在维持迁移中的作用。我们的结果表明，缺氧类器官模型早在第一天就表现出部分 EMT 特征，这些特征作为这些非迁移类器官得以保留向迁移表型的转变。在两年的延长期内，我们将继续 1) 优化我们的 DCIS 模型，纳入导管 DCIS 微环境的结构和其他组件； 2）测试连接肿瘤内在的新机制缺氧、部分/混合EMT和集体迁移； 3）抑制信号机制以阻止出现迁移表型。拟议工作的成功完成将为该领域的两个基本问题提供答案。浸润性乳腺癌的进展： 1) 是什么导致一些 DCIS 细胞迁移并发展为侵袭性肿瘤？ 2）迁移表型（IDC）如何以及在何处出现？机械论从这些研究中获得的理解将改善诊断，导致治疗策略的制定在癌前阶段阻止侵袭，从而防止患者过度治疗。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Identifying Molecular Signatures of Distinct Modes of Collective Migration in Response to the Microenvironment Using Three-Dimensional Breast Cancer Models.

使用三维乳腺癌模型识别响应微环境的集体迁移的不同模式的分子特征。

DOI：
发表时间：
2021-03-20
期刊：
影响因子：
5.2
作者：
Ardila, Diana Catalina;Aggarwal, Vaishali;Singh, Manjulata;Chattopadhyay, Ansuman;Chaparala, Srilakshmi;Sant, Shilpa
通讯作者：
Sant, Shilpa

Interplay between tumor microenvironment and partial EMT as the driver of tumor progression.

肿瘤微环境和部分EMT之间的相互作用作为肿瘤进展的驱动力。

DOI：
发表时间：
2021-02-19
期刊：
影响因子：
5.8
作者：
Aggarwal, Vaishali;Montoya, Catalina Ardila;Donnenberg, Vera S;Sant, Shilpa
通讯作者：
Sant, Shilpa

P4HA2: A link between tumor-intrinsic hypoxia, partial EMT and collective migration.

P4HA2：肿瘤内在缺氧、部分 EMT 和集体迁移之间的联系。

DOI：
发表时间：
2022-10
期刊：
影响因子：
0
作者：
Aggarwal, Vaishali;Sahoo, Sarthak;Donnenberg, Vera S;Chakraborty, Priyanka;Jolly, Mohit Kumar;Sant, Shilpa
通讯作者：
Sant, Shilpa

Mathematical Modeling of Plasticity and Heterogeneity in EMT.

EMT 中可塑性和异质性的数学模型。