A Role for KSHV in the Pathogenesis of Malignancies

KSHV 在恶性肿瘤发病机制中的作用

• 批准号: 10487195
• 负责人: Giovanna Tosato
• 金额: $ 64.18万
• 依托单位: DIVISION OF CLINICAL SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487195
• 关键词: AIDS related cancer; AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Adult; Antibodies; Aorta; B-Lymphocytes; CD3 Antigens; Cause of Death; Cell Differentiation process; Cell Line; Cell Lineage; Cell Proliferation; Cells; Clinical; Clinical Research; Clonality; Clone Cells; Complex; Derivation procedure; Development; Diffuse; Disease; Dorsal; Drug Combinations; Drug Targeting; Embryo; Endothelial Cells; Endothelium; Engraftment; Environment; Environmental Risk Factor; Epigenetic Process; Epstein-Barr Virus Infections; Future; Gene Expression; Gene Proteins; Genes; Goals; Greater sac of peritoneum; HIV; HIV Infections; HIV Seronegativity; Hematopoietic; Hematopoietic stem cells; Hodgkin Disease; Human; Human Herpesvirus 4; Human Herpesvirus 8; ITGAX gene; Immune; Immunity; Immunoblotting; Immunodeficient Mouse; Infection; Inflammation; Inflammatory; Interleukin-6; Kaposi Sarcoma; Laboratory Study; Lesion; Link; Liposomal Doxorubicin; Lubricants; Lymphocyte; Lymphoid; Lymphoma; Lymphoma cell; Lymphomagenesis; MS4A1 gene; Maintenance; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mature B-Lymphocyte; Measures; Mesenchymal; Mesoderm; Mesothelial Cell; Mesothelium; Molecular; Multicentric Angiofollicular Lymphoid Hyperplasia; Mus; Natural History; Outcome; Ovarian Carcinoma; PTPRC gene; Pathogenesis; Patients; Peritoneal; Peritoneum; Phenotype; Pilot Projects; Play; Population; Process; Research; Role; STAT3 gene; Signal Transduction; Source; Structure; Surface; Syndrome; T-Lymphocyte; Therapeutic; Tumor Angiogenesis; Tumorigenicity; Umbilical Cord Blood; VEGFA gene; Viral; Virus Diseases; Work; Yolk Sac; angiogenesis; bevacizumab; body cavity; cancer type; caveolin 1; cell growth; cell type; cytokine; effusion; epigenetic silencing; hematopoietic stem cell emergence; insight; interest; monolayer; mouse development; mouse model; neoplastic cell; novel; patient population; peripheral blood; phenotypic biomarker; prevent; primary effusion lymphoma; repaired; research clinical testing; success; transdifferentiation; tumor; tumor progression; tumorigenesis; tumorigenic; virus related cancer

We have focused primarily on the study of primary effusion lymphoma in patients with AIDS. Primary effusion lymphoma (PEL) is a Kaposi's sarcoma herpes virus (KSHV)-induced lymphoma that typically arises in body cavities of HIV-infected patients. PEL cells are often co-infected with Epstein-Barr virus (EBV). "PEL-like" lymphoma is a KSHV-unrelated lymphoma that arises in body cavities of HIV-negative patients. "PEL-like lymphoma" is sometimes EBV-positive. The derivation of PEL/"PEL-like" cells is unclear. To study PEL pathogenesis, mesothelial cells were cultured from body cavity effusions of 23 patients. These included patients with AIDS with or without PEL presenting with effusions in body cavities, and patients with ovarian carcinoma with malignant or not malignant effusions. Cell proliferation, cytokine secretion, marker phenotypes, KSHV/EBV infection and clonality were evaluated. Gene expression was measured by qPCR and immunoblotting. A mouse model of PEL was used to evaluate tumorigenicity. We found that mesothelia derived from 6 effusions of HIV-infected patients with PEL or other KSHV-associated diseases contain rare KSHV+ or EBV+ mesothelial cells. After extended culture (16-17 weeks), some mesothelial cells underwent a trans-differentiation process generating lymphoid-type CD45+/B220+, CD5+, CD27+, CD43+, CD11c+ and CD3- cells resembling "B1-cells", most commonly found in mouse body cavities. These "B1-like" cells were short-lived. However, long-term KSHV+EBV- and EBV+KSHV- clonal cell lines emerged from mesothelial cultures from two patients, which were clonally distinct from the monoclonal or polyclonal B-cell populations found in the patient's original effusion. The current study provides a novel and unifying insight into PEL and "PEL-like" lymphomagenesis. Three observations emerged from these studies. First, we discovered that mesothelial cell monolayers undergo a "mesothelial-to-lymphoid" transition (MLT)" resulting in the emergence of "lymphoid-type" cells. This discovery extends the spectrum of mesothelial functional capabilities, beyond secretion of lubricants, maintenance of surface integrity and ability to repair. MLT resembles the emergence of hematopoietic cells from the endothelium of the yolk sac and the dorsal aorta identified as "endothelial-to-hematopoietic transition" (EHT). Endothelial and mesothelial cells can undergo phenotypic and functional change through "endothelial-to-mesenchymal" (EMT) and "mesothelial-to-mesenchymal" transitions (MMT). The second discovery we made is that the lymphocytes emerging from mesothelial cultures have a "B1-like" phenotype, supporting a mesothelial origin of human "B1-type" cells. B1 lymphocytes are the main B-cell population in murine body cavities but are rare elsewhere. Human "B1-like" lymphocytes have been identified in cord and peripheral blood, but to our knowledge not in body cavities. Despite their importance in immune defense, the origin of "B1-type" cells is unclear. During mouse development, B1 lymphocytes are first detected at embryonic day (E) 8.0-8.5 in the para-aortic mesoderm prior to the emergence of hematopoietic stem cells (HSC) from the dorsal aorta, suggesting an HSC-independent origin of B1 cells. All mesothelia that line body cavities derive from the para-aortic splanchnopleural mesoderm. Thus, "B1-type" cells and mesothelia have a common developmental derivation, raising the possibility that persistence of mesodermal precursors within adult mesothelia confers B1-cell differentiation potential to these mesothelia. The third observation we made is that mesothelial cells can be infected by KSHV and EBV. The emergence of monoclonal B-lineage cell lines from mesothelial cultures suggests that PEL and PEL-like lymphoma may derive from KSHV or EBV-infected mesothelial cells. It is noteworthy that two of the 3 lines so derived were clonally distinguishable from the cells originally found in the patient. It remains possible that these lines may have emerged as a result of outgrowth from rare clones of PEL or EBV-infected cells in the original effusion. However, the evidence of MLT transition and clonal analysis suggest that a more likely explanation is that the unique clonal lines derived from the KSHV or EBV-infected mesothelial cells. Interestingly, the "indeterminate" surface phenotype of PEL cells and cell lines, including the KSHV+ 81 lines, resembles the predominant surface phenotype of the "B1-like" cells. In addition, the monoclonal KSHV-/EBV+ clone resembles the KSHV- "PEL-like" lymphoma in showing a mature B-cell phenotype and EBV infection, raising the possibility that the CD20+ cell subset recovered from mesothelial cultures is a source of "PEL-like lymphoma". From this perspective, body cavity-associated PEL and "PEL-like" malignancies would have a common mesothelial derivation. Ongoing and future studies will be focused on determining if this new understanding of PEL pathogenesis can be exploited to prevent PEL development and treat it by targeting the mesothelium. To pursue this goal, we are developing a mouse model in which cultures of human mesothelium infected or not infected with KSHV are explanted into immunodeficient mice. We want to observe the localization of peritoneal explants of mesothelium, its natural history in the mouse body cavity, the potential emergence of "B1"-type cells and the potential emergence of PEL. Since inflammation within the body cavities is commonly detected in patients with AIDS, we will explore the role of experimental inflammation in the emergence of human B1-type cells and PEL. We have obtained engraftment of PEL-derived primary mesothelial cells expanded in culture and transferred into the peritoneal cavity of immunodeficient mice (NSG mice). Macroscopic examination of the peritoneal cavity showed the presence of focal nodular structures and more diffuse thickening of the peritoneum. We did not detect the presence of PEL-type cells floating in the peritoneal cavity. We are currently further evaluating these lesions and their relationship with KSHV diseases. Following up on our previous laboratory studies highlighting the role of VEGFA in the pathogenesis of KS, we have collaborated with the clinical group of HAMB in a pilot study of the anti-VEGF antibody Bevazizumab combined with liposomal doxorubicin in a group of patients with advanced KS. Evaluation of this clinical study showed that patients with advanced KS are responsive to the combination of liposomal doxorubicin and Bevazizumab. Future studies will need to directly assess if the drug combination yields a better outcome than liposomal doxorubicin alone in patients with advanced KS. In a collaborative study, we have examined further the role of viral IL-6 in HSHV infection. We have now discovered that vIL-6 epigenetically silences caveolin-1 (CAV1) expression to promote angiogenesis and tumorigenesis by regulating the formation of STAT3-DNMT1 complex. This new information will guide future studies on epigenetic changed induced by KHSV infection of endothelial cells.

我们主要关注艾滋病患者的原发性积液淋巴瘤的研究。原发性淋巴瘤（PEL）是Kaposi的肉瘤疱疹病毒（KSHV）诱导的淋巴瘤，通常在受HIV感染的患者的体腔中产生。 PEL细胞通常与Epstein-Barr病毒（EBV）共感染。 “ PEL样”淋巴瘤是一种KSHV无关的淋巴瘤，在HIV阴性患者的体腔中产生。 “ PEL样淋巴瘤”有时是EBV阳性的。 pel/“ pel样”细胞的推导尚不清楚。为了研究PEL发病机理，从23例患者的体腔积液中培养间皮细胞。其中包括患有或没有PEL的AIDS患者，在体腔中呈液体，以及患有恶性或不恶性积液的卵巢癌患者。评估了细胞增殖，细胞因子分泌，标记表型，KSHV/EBV感染和克隆性。通过qPCR和免疫印迹测量基因表达。 PEL的小鼠模型用于评估肿瘤性。我们发现，间皮源自6种HIV感染的PEL或其他KSHV相关疾病的患者含有罕见的KSHV+或EBV+间皮细胞。在扩展培养（16-17周）之后，一些间皮细胞进行了产生淋巴样型CD45+/B220+，CD5+，CD27+，CD43+，CD43+，CD11C+和CD3-细胞类似于“ B1-Cells”的反差异过程。这些“ B1状”细胞是短暂的。然而，长期KSHV+EBV和EBV+KSHV-克隆细胞系来自来自两名患者的间皮培养物，这些患者与患者原始积液中发现的单克隆或多克隆B细胞群体不同。当前的研究提供了对PEL和“ PEL样”淋巴细胞造成的新颖而统一的见解。这些研究得出了三个观察结果。首先，我们发现，间皮细胞单层会经历“间皮上到淋巴”“过渡（MLT）”，导致“淋巴样型”细胞的出现。这一发现扩展了间皮功能的频谱，从而扩大了间皮功能的频谱，超越了润滑体的分泌，超越了润滑性的分泌，可修复水平的分泌。蛋黄囊的内皮和背主动脉被确定为“内皮到山地组织的过渡”（EHT）。来自间皮培养的淋巴细胞具有“ B1”表型，支持人类“ B1型”细胞的介体起源，尽管他们的B1型“ B1样”是少量的，但在其他地方却差不多在免疫防御中的重要性，在小鼠发育过程中，“ B1型”细胞的起源尚不清楚。腔体从para-Apara-splanchnoplearal的中胚层衍生出来来自间皮培养的单克隆B线细胞系的出现表明，PEL和PEL样淋巴瘤可能衍生自KSHV或EBV感染的间皮细胞，值得注意的是，这三种细胞中的两条线可能与这些细胞相同，因此可能会发现这些细胞。然而，在原始积液中，EBV感染的细胞是MLT过渡和克隆分析的证据表明，更有可能的解释是从KSHV或EBV感染的中心细胞中得出的独特克隆线此外，单克隆KSHV-/EBV+克隆在显示成熟的B细胞表型和EBV感染中类似于KSHV-“ PEL”淋巴瘤，从而增加了CD20+细胞子群的可能性，即从中皮培养物中恢复了从pel样淋巴瘤中的peel-perceptive cavity，pel-pece and pel and pel and pel。具有持续的间皮性衍生物，将来将利用这种对PEL发病机理的新理解来防止PEL发育，并通过靶向中皮来追求这一目标，我们正在开发一个小鼠模型。在艾滋病患者中，在体内的潜在出现，在艾滋病患者中，它通常会检测到中皮的炎症，从而探索了pel的潜在出现。免疫缺陷小鼠的腹膜（NSG小鼠）的宏观检查表明，局部结构的存在和腹膜的弥漫性增厚我们目前没有在腹膜上进行研究。为了强调VEGFA在KS的发病机理中的作用，我们与汉堡的临床组合作，在一项抗VEGF抗体贝韦齐珠单抗的试点研究中，与脂质体阿霉素相结合。未来的研究将需要直接评估与脂质体阿霉素相比，在一项协作研究中，单独的脂质体阿霉素。 STAT3-DNMT1复合物。