Inflammatory Breast Cancer: Factors Contributing to Dissemination

描述（由申请人提供）：IBC是原发性乳腺癌的最致命形式，不成比例地针对年轻女性。IBC占美国所有乳腺癌病例的<5%，但其在非洲裔美国妇女中的发病率明显较高（10.1%）。据报道，在埃及，IBC的发病率占所有乳腺癌病例的5- 10%，并且正在上升。广泛的真皮淋巴浸润和其中的肿瘤栓塞导致淋巴阻塞，这是疾病的炎症性质的基础。我们已经表明巨噬细胞浸润IBC微环境，包围IBC栓子，并存在于IBC患者的腋血中。我们的工作假设是，IBC相关的巨噬细胞在肿瘤栓塞的形成和癌细胞在肿瘤中的扩散中起着至关重要的作用，这是IBC的两个表型标志。该FIRCA提案的具体目的是：1）建立IBC细胞的3D培养物和a）评估侵袭性表型; B）确定推定的IBC标志物蛋白的表达/定位; c）定量条件培养基中细胞因子、趋化因子和生长因子的表达;和d）半胱氨酸组织蛋白酶的表达/分泌/活性; 2）表征来自IBC和非IBC患者的肿瘤栓子、肿瘤、阳性淋巴结、腋窝支流血液和外周血中的单核细胞/巨噬细胞表型和含量; 3）从IBC腋支血和外周血中分离单核细胞，a）定量分泌的细胞因子、趋化因子和生长因子的表达，和B）半胱氨酸组织蛋白酶的表达/分泌/活性; 5）建立从IBC腋支血和外周血分离的IBC细胞和单核细胞的3D共培养物，和a）评估侵袭性表型; B）测定推定IBC标记蛋白的表达/定位; c）定量条件培养基中细胞因子、趋化因子和生长因子的表达;和d）半胱氨酸组织蛋白酶的表达/分泌/活性;和e）对IBC样本中鉴定的细胞表面受体进行免疫染色共培养物。未来的方向将使用目标1-5中获得的信息来选择半胱氨酸组织蛋白酶、细胞因子、趋化因子和生长因子，它们可能被靶向以减少IBC的侵袭性表型。IBC在埃及的积极表现意味着迫切需要制定研究计划，以立即帮助乳腺癌患者。我们预计，我们的研究结果将有助于表征IBC的病理生物学基础，这是优先考虑治疗干预的新靶点的先决条件。这项研究将主要在埃及开罗大学与莫纳穆斯塔法·穆罕默德合作进行，作为NIH批准号R 01 CA 131990（2008年8月1日至2013年5月31日）的扩展。 公共卫生相关性：该提案汇集了埃及和美国研究人员的多学科团队，共同努力，最终目标是确定可用于针对IBC的新疗法的药物途径，IBC是一种高度致命的癌症，在年轻女性中发现不成比例，在埃及发病率很高。本申请中提出的研究的成功完成将产生经验证的体外模型系统，其可用于筛选新疗法的临床前研究，并且还可鉴定IBC的生物标志物。我们的目的是提高诊断和最终治疗管理这种毁灭性的癌症。

DESCRIPTION (provided by applicant): IBC is the most lethal form of primary breast cancer and disproportionately targets younger women. IBC accounts for <5% of all breast cancer cases in the United States, but its incidence is significantly higher among African-American women (10.1%). In Egypt, the incidence of IBC is reported to be from 5- 10% of all breast cancer cases and is on the rise. Extensive dermal lymphatic invasion and tumor emboli therein result in lymphatic obstruction that underlies the inflammatory nature of the disease. We have shown that macrophages infiltrate the IBC microenvironment, surround IBC emboli and are present in axillary blood of IBC patients. Our working hypothesis in this proposal is that IBC-associated macrophages play a crucial role in the formation of tumor emboli and dissemination of the cancer cells in the lymphatics, two of the phenotypic hallmarks of IBC. The Specific Aims of this FIRCA proposal are to: 1) Establish 3D cultures of IBC cells and a) assess invasive phenotype; b) determine expression/localization of putative IBC marker proteins; c) quantify expression of cytokines, chemokines and growth factors in conditioned media; and d) expression/secretion/activity of cysteine cathepsins; 2) Characterize monocyte/macrophage phenotype and content in tumor emboli, tumors, positive lymph nodes, axillary tributary blood and peripheral blood from IBC and non-IBC patients; 3) Isolate monocytes from IBC axillary tributary blood and peripheral blood and a) quantify expression of secreted cytokines, chemokines and growth factors, and b) expression/secretion/activity of cysteine cathepsins; 4) Immunostain paraffin blocks of IBC specimens for cell surface receptors of selected cytokines, chemokines and growth factors; and, 5) Establish 3D cocultures of IBC cells and monocytes isolated from IBC axillary tributary blood and peripheral blood and a) assess invasive phenotype; b) determine expression/localization of putative IBC marker proteins; c) quantify expression of cytokines, chemokines and growth factors in conditioned media; and d) expression/secretion/activity of cysteine cathepsins; and, e) immunostain cocultures for cell surface receptors identified in IBC specimens. Future directions will use the information obtained in Aims 1-5 to select cysteine cathepsins, cytokines, chemokines and growth factors that might be targeted to reduce the invasive phenotype of IBC. The aggressive manifestation of IBC in Egypt means that there is a pressing need to develop research programs that can be of immediate assistance to patients suffering from breast cancer. We anticipate that our results will contribute to the characterization of the pathobiological underpinnings of IBC, a prerequisite for prioritizing novel targets for therapeutic intervention. This research will be done primarily in Egypt at Cairo University in collaboration with Mona Mostafa Mohamed, as an extension of NIH Grant No. R01CA131990, 8-01-2008 to 5-31-2013. PUBLIC HEALTH RELEVANCE: This proposal brings together a multi-disciplinary team of Egyptian and American investigators in a collaborative effort with the ultimate goal of identifying druggable pathways that can be targeted for new therapies against IBC, a highly lethal cancer that is found disproportionately in young women and has a high incidence in Egypt. Successful completion of the studies proposed in this application will result in validated in vitro model systems that can be used for preclinical studies screening new therapeutics and may also identify biomarkers for IBC. Our intent is to improve the diagnosis and eventually the therapeutic management of this devastating cancer.