Inflammatory Breast Cancer: Factors Contributing to Dissemination

描述（由申请人提供）：IBC是最致命的原发性乳腺癌，主要针对年轻女性。在美国，IBC占所有乳腺癌病例的不到5%，但其在非裔美国女性中的发病率明显更高（10.1%）。据报道，在埃及，IBC的发病率占所有乳腺癌病例的5- 10%，并且呈上升趋势。广泛的真皮淋巴浸润和肿瘤栓塞导致淋巴阻塞，这是炎症性疾病的基础。我们发现巨噬细胞浸润IBC微环境，包围IBC栓子，并存在于IBC患者的腋窝血中。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淋巴管中癌细胞的传播中起着至关重要的作用，这是IBC的两个表型特征。FIRCA提案的具体目的是：1)建立IBC细胞的3D培养和a)评估侵袭性表型；b)确定IBC标记蛋白的表达/定位；C)在条件培养基中量化细胞因子、趋化因子和生长因子的表达；d)半胱氨酸组织蛋白酶的表达/分泌/活性；2)对IBC和非IBC患者的肿瘤栓子、肿瘤、阳性淋巴结、腋窝支血和外周血中单核/巨噬细胞的表型和含量进行表征；3)从IBC腋窝支血和外周血中分离单核细胞，a)量化分泌细胞因子、趋化因子和生长因子的表达，b)半胱氨酸组织蛋白酶的表达/分泌/活性；4)对IBC标本石蜡块进行免疫染色，筛选细胞因子、趋化因子和生长因子的细胞表面受体；5)建立IBC细胞和从IBC腋窝分支血和外周血中分离的单核细胞的三维共培养，a)评估侵袭性表型；b)确定IBC标记蛋白的表达/定位；C)在条件培养基中量化细胞因子、趋化因子和生长因子的表达；d)半胱氨酸组织蛋白酶的表达/分泌/活性；e)在IBC标本中鉴定的细胞表面受体的免疫染色共培养。未来的方向将利用Aims 1-5中获得的信息，选择可能靶向降低IBC侵袭性表型的半胱氨酸组织蛋白酶、细胞因子、趋化因子和生长因子。IBC在埃及的侵略性表现意味着迫切需要制定研究计划，可以立即帮助乳腺癌患者。我们预计我们的结果将有助于表征IBC的病理生物学基础，这是优先考虑治疗干预新靶点的先决条件。这项研究将主要在埃及的开罗大学与Mona Mostafa Mohamed合作进行，作为NIH拨款号的延伸。R01CA131990, 8-01-2008至5-31-2013。

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.