Molecular Mechanisms Of Growth Control And Carcinogenesis

生长控制和致癌的分子机制

• 批准号: 8743736
• 负责人: J Gutkind
• 金额: $ 161.15万
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 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/8743736
• 关键词: Acquired Immunodeficiency Syndrome; Adhesions; Adult; Agonist; Antineoplastic Agents; Behavior; Binding; Blue Nevus; Cell Nucleus; Cell Proliferation; Cells; Colon Carcinoma; Coupled; Cutaneous Melanoma; Cytokine Activation; Development; Dissection; Down-Regulation; Drosophila genus; Employee Strikes; Endocrine Gland Neoplasms; Endometrial; Endothelial Cells; Eukaryotic Initiation Factor-4E; Event; Exhibits; FOS gene; Family; Family member; Frequencies; G Protein-Coupled Receptor Genes; G-Protein-Coupled Receptors; G-substrate; GNAQ gene; GTP-Binding Proteins; Genes; Genetic; Genetic Transcription; Genetic Translation; Genetically Engineered Mouse; Glutamate Receptor; Growth; Growth Factor; Growth Factor Receptors; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Head and Neck Cancer; Head and Neck Squamous Cell Carcinoma; Human; Human Herpesvirus 8; Hydrolysis; In Vitro; Inflammation; JUN gene; Kaposi Sarcoma; Lesion; Ligands; Link; Lymphangiogenesis; Lymphatic Endothelial Cells; Lysophosphatidic Acid Receptors; MAPK14 gene; Malignant - descriptor; Malignant Neoplasms; Mammalian Cell; Marketing; Mediating; Melanoma Cell; Meninges; Messenger RNA; Metastasis Suppressor Genes; Mitogens; Modeling; Molecular; Molecular Target; Mus; Mutation; N-terminal; Normal Cell; Ocular Melanoma; Oncogenes; Ovarian; Parathyroid gland; Pathologic Neovascularization; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Pituitary Neoplasms; Primary carcinoma of the liver cells; Process; Proteins; RNA Interference; Reporter; Reporting; Role; Second Messenger Systems; Semaphorins; Signal Transduction; Sirolimus; System; Thyrotropin Receptor; Transcription Factor AP-1; Translations; Uveal Melanoma; Vascular Endothelial Growth Factor C; Viral; Work; Xenograft procedure; angiogenesis; base; biliary tract; cancer cell; cancer prevention; cancer therapy; carcinogenesis; cell growth; cell transformation; gain of function mutation; genome-wide; growth promoting activity; human FRAP1 protein; human SMO protein; in vivo; in vivo Model; mTOR Inhibitor; mTOR inhibition; member; mutant; neoplastic; novel; novel strategies; overexpression; pancreatic neoplasm; paracrine; plexin; pre-clinical; prevent; protein protein interaction; receptor; receptor coupling; response; rho; rho GTP-Binding Proteins; second messenger; sphingosine 1-phosphate; thyroid neoplasm; tumor; tumor progression; tumorigenesis

50% Effort. Molecular dissection of the pathway linking growth factor receptors to the nucleus: their role in normal cell growth and cancer. The emerging mutational landscape of G-proteins and G-protein coupled receptors in cancer. The presence of genetic alterations in G proteins and GPCRs were initially restricted to only few neoplastic lesions in endocrine tumors. However, we have recently reported the widespread presence and high frequency of mutations in GPCRs and G proteins in most tumor types. Specifically, in a recent in depth analysis of the human oncogenome we have found that a striking 4.2% of all tumor sequences exhibit activating mutations in GNAS (encoding Gs), including thyroid and pituitary tumors, as well as colon cancer, hepatocellular carcinoma, and parathyroid, ovarian, endometrial, biliary tract, and pancreatic tumors. Mutually exclusive activating mutations in GNAQ or GNA11 (encoding Gq family members) occur in 5.6% of tumors, including >66% of ocular melanomas, thus providing a clear example of a human malignancy that is initiated by gain of function mutations in Gq and G11 proteins. GNAQ and GNA11 mutations are also found in tumors arising from the meninges (59%), in most blue nevi of the skin (83%), and in a subset of cutaneous melanomas (6%). Surprisingly, nearly 20% of human cancers harbor mutations in GPCRs, including frequent mutations in thyroid-stimulating hormone receptor (TSHR), smoothened (SMO), glutamate receptors (GRMs), members of the adhesion family of GPCRs, and receptors for lysophosphatidic acid (LPA) and sphingosine 1 phosphate (S1P). Overall, as GPCRs are the target of >25% drugs in the market, we expect that this information can be exploited for the development of novel strategies targeting GPCRs, G proteins, or their aberrant signaling circuitry for cancer prevention and treatment. A genome-wide RNAi screen reveals a Trio-regulated Rho GTPase network mediating GPCR-initiated mitogenic signaling and cancer growth. Multiple mitogens stimulate Gq-coupled GPCRs, and this receptor family contributes to cancer growth in many human malignancies. We have used Gq-coupled receptors activated solely by synthetic ligands (RASSLs) to build, and hence understand, GPCR-regulated signaling networks in normal and cancer cells. Gq-RASSL transduces potent mitogenic signals and is transforming if persistently activated, a process that requires the expression of c-Jun and c-Fos AP-1 family members. To investigate how GPCRs regulate AP-1-dependent gene transcription, we performed a genome-wide high-throughput RNAi screen in Drosophila S2 cells expressing Gq-GPCR and an AP-1 reporter system. Interestingly, we found that Rho family GTPases, specifically Rho and Rac, and their downstream effectors, such as Pak and Jun N-terminal kinase (JNK) were integral to AP-1 activation. While molecules linking GPCRs to the hydrolysis of phosphatidylinositol and PKC activation were dispensable, we found that Trio, a Rho guanine nucleotide exchange factor (GEF) that binds directly to Gq, is essential for AP-1 activation by Gq-coupled GPCRs. Remarkably, Trio is essential for the activation of JNK and p38 MAPKs, c-Jun and c-Fos expression, AP1 activation, and cell proliferation and transformation in mammalian cells. Furthermore, many cancers, including head and neck cancer (HNSCC) overexpress Trio due to genetic amplification. Trio downregulation prevents the mitogenic response to Gq-coupled receptor agonists in normal and cancer cells, and halts the growth of uveal melanoma cells harboring Gq mutations. These findings indicate that the growth promoting activity of Gq-linked GPCRs involves the activation of AP-1 by a Rho-GTPase network through Trio, and that this process is governed by highly specific protein-protein interactions and phosphorylation events rather than by diffusible second messengers. 30% Effort. Molecular basis of developmental and tumor-induced angiogenesis. Molecular mechanisms by which Semaphorins and Plexins control angiogenesis and lymphangiogenesis: Semaphorin 3E (Sema3E) and its receptor Plexin-D1 control the patterning of the developing vasculature. However, it was not known whether Sema3E-Plexin-D1 signals in adult and in pathological angiogenesis. We have recently observed that Sema3E behaves as a potent natural anti-lymphangiogenic molecule in a number of in vivo models of developmental and tumor-induced lymphangiogenesis. In particular, we observed that Sema3E provokes the rapid retraction of lymphatic endothelial cells, and diminish the pro-angiogenic activity of VEGF-C and S1P in vitro and in vivo, and prevents the pro-lymphangiogenic effect of HNSCC cells when grown in mouse xenografts. We are now investigating the underlying molecular mechanisms by which Sema3E acts in lymphatic endothelial cells. We have also focused in the possibility that another semaphorin, Sema3F, may represent an anti-lymphangiogenic metastasis suppressor gene, given that one of its co-receptors, NRP2, is expressed primarily in lymphatic endothelial cells, and that Sema3F gene loss is a frequent event in advanced HNSCC lesions. 20% effort. AIDS-associated Kaposis sarcoma: molecular mechanisms. Dephosphorylated 4EBP disrupts paracrine transformation by the KSHV vGPCR oncogene upon mTOR inhibition: Early work from our group led to the identification of the Akt/mTOR pathway as a critical signaling axis contributing to KSHV-induced cancer progression, and treatment of KS patients with rapamycin provided the first evidence of the antineoplastic activity of mTOR inhibitors in humans. Thus, the study of KS may provide a unique opportunity to dissect the contribution of specific mTOR substrates to cancer development. We focused on a direct target of mTOR, 4EBP1/2/3 (4EBP), which inhibits the translation of eukaryotic initiation factor 4E (4E)-bound mRNAs. 4EBP phosphorylation by mTOR relieves its inhibitory activity, hence resulting in increased 4E-dependent mRNA translation. We developed a paracrine transformation model, recapitulating the cellular composition of KS lesions, in which vGPCR-expressing cells promote the rapid proliferation of endothelial cells expressing KSHV-latent genes by the release of growth factors. Using this model, we showed that the accumulation of dephosphorylated 4EBP in response to rapamycin or by the expression of an mTOR-insensitive mutant of 4EBP1 is sufficient to disrupt the 4E function downstream of mTOR thereby halting KS development. These findings may provide a preclinical platform and the rationale for the development of novel mTOR inhibiting agents that may selectively disrupt the mTOR-4EBP interaction for the treatment of KS and other tumor lesions exhibiting hyperactive mTOR pathway The IKK kinase is critical for vGPCR-induced tumorigenesis. In previous studies we have shown that vGPCR stimulates the expression of pro-angiogenic cytokines by the activation of NFB. We have recently observed that this process involves the increased activity of IKK, an IKK-related kinase, and that IKK is critically required for vGPCR tumorigenesis. Loss of IKK in genetically engineered mice effectively abrogated NFB activation and tumorigenesis triggered by vGPCR. Collectively, our findings uncover the critical role of IKK in promoting inflammation and tumorigenesis induced by a viral GPCR.

50％的努力。将生长因子受体与细胞核联系起来的途径的分子解剖：它们在正常细胞生长和癌症中的作用。 癌症中G蛋白和G蛋白偶联受体的新兴突变景观。 G蛋白和GPCR中遗传改变的存在最初仅限于内分泌肿瘤中的少量肿瘤病变。但是，我们最近报道了大多数肿瘤类型中GPCR和G蛋白中突变的广泛存在和高频。具体而言，在对人类癌基因组的最新深度分析中，我们发现所有肿瘤序列中有4.2％的4.2％在GNA（编码GS）中表现出激活突变，包括甲状腺和垂体肿瘤，以及结肠癌，肝细胞癌癌和肝癌，以及结肠癌甲状旁腺，卵巢，子宫内膜，胆道和胰腺肿瘤。在GNAQ或GNA11中相互排斥的激活突变（编码GQ家族成员）发生在5.6％的肿瘤中，包括> 66％的眼部黑色素瘤，因此为人类恶性肿瘤提供了一个明确的例子，该肿瘤是由GQ和G11中功能突变增益引发的。蛋白质。在脑膜（59％），皮肤的大多数蓝色NEVI（83％）以及皮肤黑色素瘤（6％）的一部分中，GNAQ和GNA11突变也存在于脑膜（59％）的肿瘤中（59％）。令人惊讶的是，将近20％的人类癌症藏有GPCR中的突变，包括甲状腺刺激激素受体（TSHR）中的频繁突变（TSHR），平滑（SMO），谷氨酸受体（GRMS），GPCR的粘附家族的成员以及溶质磷酸磷酸磷酸化的受体的成员（LPA）和1磷酸盐（S1P）。总体而言，由于GPCR是市场上> 25％药物的靶标，因此我们希望可以利用此信息来开发针对GPCR，G蛋白的新型策略，或者以进行癌症预防和治疗的异常信号通路。 全基因组的RNAi筛查揭示了三个调节的Rho GTPase网络，介导GPCR引起的有丝分裂信号传导和癌症生长。多种有丝分裂剂刺激GQ耦合的GPCR，该受体家族在许多人类恶性肿瘤中有助于癌症的生长。我们已经使用仅由合成配体（RASSL）激活的GQ耦合受体来构建，因此了解正常和癌细胞中GPCR调节的信号网络。 GQ-RASSL会传递有效的有丝分裂信号，并在持续激活（需要C-Jun和C-Fos AP-1家族成员表达的过程）中转换。为了研究GPCR如何调节AP-1依赖性基因转录，我们在表达GQ-GPCR和AP-1报告基因系统的果蝇S2细胞中进行了全基因组的高通量RNAi筛选。 有趣的是，我们发现Rho家族GTPases，特别是RHO和RAC及其下游效应子，例如PAK和JUN N末端激酶（JNK）是AP-1激活不可或缺的。虽然将GPCR连接到磷脂酰肌醇和PKC激活的水解的分子是可分配的，但我们发现Trio是直接与GQ结合的Rho Guanine核苷酸交换因子（GEF），对于通过GQ耦合GPCRS激活AP-1是必不可少的。值得注意的是，三重奏对于JNK和p38 MAPK，C-JUN和C-FOS表达，AP1激活以及细胞增殖和转化是必不可少的。此外，由于遗传扩增，许多癌症，包括头颈癌（HNSCC）过表达三重奏。三重点下调可防止正常和癌细胞中对GQ偶联受体激动剂的有丝分裂反应，并阻止了具有GQ突变的卵巢黑色素瘤细胞的生长。这些发现表明，促进GQ连锁GPCR的生长活性涉及通过三重奏通过Rho-GTPase网络激活AP-1，并且该过程受高度特异性的蛋白质蛋白质相互作用和磷酸化事件的控制，而不是通过可扩散的第二个。使者。 30％的努力。发育和肿瘤诱导的血管生成的分子基础。 词素和斑点蛋白控制血管生成和淋巴管生成的分子机制：Semaphorin 3E（SEMA3E）及其受体plexin-D1控制发育中的脉管系统的模式。然而，尚不知道成人和病理血管生成中的Sema3e-plexin-d1信号。我们最近观察到，SemA3E在许多具有发育和肿瘤诱导的淋巴管生成的体内模型中是有效的天然抗淋巴管分子。特别是，我们观察到SEMA3E会引起淋巴内皮细胞的快速缩回，并在体外和体内降低VEGF-C和S1P的促血管生成活性，并预防小鼠Xenogografts中HNSCC细胞的促淋巴结效应。我们现在正在研究SEMA3E在淋巴内皮细胞中作用的潜在分子机制。我们还集中于这样的可能性：鉴于其一个共受体NRP2主要在淋巴内皮细胞中表达，并且该SEMA3F基因损失频繁表达，鉴于其一个共受体NRP2可能是一种抗淋巴染转移抑制基因的可能性，并且高级HNSCC病变中的事件。 20％的努力。与艾滋病相关的kaposis肉瘤：分子机制。 Dephosphory的4EBP在MTOR抑制后通过KSHV VGPCR癌症破坏了旁分泌的转化：我们组的早期工作导致AKT/MTOR途径鉴定出AKT/MTOR途径，作为对KSHV诱导的癌症进展的关键信号轴，并鉴定MTOR抑制剂在人类中的抗肿瘤活性的第一个证据。因此，对KS的研究可能会提供独特的机会，以剖析特定MTOR底物对癌症发展的贡献。我们专注于MTOR的直接目标，即4EBP1/2/3（4EBP），该目标抑制了真核开始因子4E（4E）结合mRNA的翻译。 MTOR通过MTOR磷酸化的4EBP磷酸化减轻了其抑制活性，从而导致4E依赖性mRNA翻译增加。我们开发了一个旁分泌转化模型，概括了KS病变的细胞组成，其中表达VGPCR的细胞通过释放生长因子促进表达KSHV latent基因的内皮细胞的快速增殖。使用该模型，我们表明，对雷帕霉素的响应或4EBP1的mTOR不敏感突变体的表达足以破坏MTOR下游的4E功能，从而避免KS发育。这些发现可能提供一个临床前平台和开发新型MTOR抑制剂的基本原理，这些抑制剂可能会选择性地破坏MTOR-4EBP相互作用，用于治疗KS和其他表现出多活跃途径的KS和其他肿瘤病变 IKK激酶对于VGPCR诱导的肿瘤发生至关重要。在先前的研究中，我们表明VGPCR通过NFB的激活刺激促血管生成细胞因子的表达。我们最近观察到，这一过程涉及与IKK相关激酶IKK的活动增加，并且IKK对于VGPCR肿瘤发生了至关重要的。基因工程小鼠IKK的丧失有效地消除了NFB激活和由VGPCR触发的肿瘤发生。总体而言，我们的发现发现了IKK在促进病毒GPCR引起的炎症和肿瘤发生中的关键作用。