Ron Receptor Tyrosine Kinase Signaling in Breast Cancer

乳腺癌中的 Ron 受体酪氨酸激酶信号转导

• 批准号: 10571969
• 负责人: Susan E Waltz
• 金额: --
• 依托单位: CINCINNATI VA MEDICAL CENTER RESEARCH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10571969
• 关键词: Affect; American; Apoptosis; Biological; Blood Circulation; Breast; Breast Cancer Cell; Breast Cancer Model; Breast Epithelial Cells; CD8-Positive T-Lymphocytes; Cancer Death Rates; Cancer Etiology; Cell Count; Cell Proliferation; Cell physiology; Cells; Cessation of life; Clinical Research; Combined Modality Therapy; Cytotoxic T-Lymphocytes; Data; Death Rate; Detection; Development; Diagnosis; Disease; Early Diagnosis; Exhibits; Female; Functional disorder; Funding; Gene Expression Profile; Growth; Healthcare; Hepatocyte; Human; IRAK4 gene; Immune; Immunodeficient Mouse; Inflammation; Inflammatory; Innate Immune Response; Interferon Suppression; Interferon Type I; Interferons; Interleukin Suppression; Invaded; Knowledge; Laboratories; Ligands; Link; Macrophage; Mammary Neoplasms; Mediating; Mediator; Messenger RNA; Metastatic Neoplasm to the Liver; Metastatic Neoplasm to the Lung; Modeling; Molecular; Mouse Mammary Tumor Virus; Mus; Neoplasm Metastasis; Oncogenic; Organoids; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Phenocopy; Phenotype; Phosphotransferases; Play; Polyomaviruses Middle T Proteins; Production; Prognosis; Progressive Disease; Protein Deficiency; Proteins; Publishing; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Recurrence; Regulation; Role; Signal Pathway; Signal Transduction; T cell response; T-Lymphocyte; Testing; Therapeutic; Transgenic Mice; Transplantation; Tumor Immunity; Tumor Promotion; Up-Regulation; Veterans; Woman; Women' s Group; Women' s Health; Work; aggressive breast cancer; attenuation; breast cancer progression; breast tumorigenesis; cancer subtypes; clinically relevant; combat; combinatorial; comparison control; cytokine; efficacy evaluation; experimental study; immune cell infiltrate; immunoregulation; inhibitor; innate immune sensing; insight; knock-down; macrophage stimulating protein; malignant breast neoplasm; mammary; mammary epithelium; migration; military veteran; molecular subtypes; mortality; mouse model; neoplastic cell; novel; novel marker; overexpression; paracrine; permissiveness; pharmacologic; protein function; receptor; reconstitution; recruit; response; stem-like cell; transcription factor; transcriptome sequencing; treatment response; treatment strategy; tumor; tumor growth; tumor initiation; tumor microenvironment; tumor progression

Breast cancer (BC) is the second leading cause of cancer related deaths among women in the US. While advances in treatment and early detection have contributed to a decline in death rates, BC is still a major killer of women both in the US and globally highlighting the need to identify new molecular circuits and treatment strategies to combat this disease. Work from our funded Merit studies and others has shown that the RON receptor tyrosine kinase is highly overexpressed in human BCs and is an independent predictor of worse overall survival, early death and metastatic progression. Interestingly, our recent studies show that RON expression is independent of molecular subtype suggesting that this protein may serve as a powerful new marker for metastasis, poor prognosis and recurrence across all BCs. While RON overexpression is a decisive factor in human BC progression, our knowledge about the mechanisms by which RON promotes the development of aggressive disease is limited. Previously, our laboratory produced transgenic mice with mammary-specific RON overexpression. These mice, referred to as MMTV-RON mice, mimic the aggressive disease pattern observed in human patients with all female mice developing highly metastatic breast tumors. In this clinically relevant model, we discovered that mammary tumors upregulate the RON ligand, hepatocyte growth factor-like protein (HGFL). Utilizing MMTV-RON mice deficient in HGFL, we published the requirement of HGFL for oncogenic RON activation and mammary tumor growth and progression. Utilizing HGFL deficient BC cells, we showed that reconstitution of HGFL promoted tumor cell autonomous survival, migration and invasion. Further, we showed that global HGFL loss also reduced breast cancer stem-like cell numbers and altered the tumor microenvironment (TME) by regulating immune cell recruitment, macrophage polarization, and cytotoxic T cell responses. In complementary studies using the well-characterized polyomavirus middle T antigen mouse model of BC, we show that RON loss specifically in mammary epithelial cells phenocopies the reduced growth, metastasis and changes in the TME as global and tumor cell specific HGFL loss. We also demonstrate a coordinate upregulation of HGFL and RON in human BCs. To gain mechanistic insights into the RON signaling pathway, we performed RNA-Seq analyses of RON modulated human and murine BC cells. Surprisingly, we found RON predominately suppressed pathways involved in Type I interferon (IFN), cytokine and pattern recognition receptor (PRR) signaling. Investigating these pathways, we discovered a novel connection between RON and suppression of IRAK4 (interleukin-1 receptor-associated kinase 4), an identified mediator of innate immune responses downstream of PRRs. We also identified a reduction in IRF7, a key regulator of Type I IFNs, with RON loss and IRAK4 expression. We show that IRAK4 introduction into RON expressing cells phenocopies RON loss, while treatment of RON deficient cells with an IRAK4 inhibitor phenocopies RON expression with respect to tumor growth and Type I IFN production. Based on this data, this proposal will test the hypothesis that upregulation of tumor cell intrinsic RON promotes BC progression through effects on both the tumor proper and TME through mechanisms that involve suppression of IRAK4 and ensuing Type I IFN production. Two Aims are proposed to (i) determine the functional significance of tumor cell intrinsic RON signaling and Type I IFN disruption in establishing a tumor permissive microenvironment to support BC growth, and (ii) define the role of RON activation in suppressing the IRAK4-Type I IFN signaling axis to promote BC growth and metastasis. This work is expected to provide new insights into mechanisms underlying aggressive BCs and will be instructive in accessing combination therapies to direct clinical studies to combat BC deaths.

乳腺癌（BC）是美国女性癌症相关死亡的第二大原因。而 虽然治疗和早期发现的进步有助于降低死亡率，但BC仍然是一个主要杀手 在美国和全球范围内的妇女强调需要确定新的分子电路和治疗 应对这种疾病的策略。我们资助的Merit研究和其他研究表明，罗恩 受体酪氨酸激酶在人类BC中高度过表达，是整体恶化的独立预测因子。 存活、早期死亡和转移进展。有趣的是，我们最近的研究表明，罗恩表达是 与分子亚型无关，这表明该蛋白可能作为一种强有力的新标记物， 转移、预后差和复发。虽然罗恩过表达是一个决定性因素， 人类BC进展，我们对罗恩促进BC发展的机制的了解， 侵袭性疾病是有限的。在此之前，我们的实验室生产了具有乳腺特异性罗恩的转基因小鼠 过度表达这些小鼠，称为MMTV-RON小鼠，模仿观察到的侵袭性疾病模式 在人类患者中，所有雌性小鼠都患有高转移性乳腺肿瘤。在这个临床相关的 模型，我们发现乳腺肿瘤上调罗恩配体，肝细胞生长因子样蛋白 （HGFL）。利用HGFL缺陷的MMTV-RON小鼠，我们发表了HGFL对致癌性的要求。 罗恩活化与乳腺肿瘤生长和进展。利用HGFL缺陷型BC细胞，我们表明， HGFL的重建促进了肿瘤细胞的自主存活、迁移和侵袭。此外，我们还展示了 全球HGFL缺失也减少了乳腺癌干细胞样细胞的数量， 微环境（TME）通过调节免疫细胞募集、巨噬细胞极化和细胞毒性T细胞 应答在使用充分表征的多瘤病毒中T抗原小鼠模型的补充研究中， 我们发现，乳腺上皮细胞中罗恩的损失特异性地表现为生长减少， 转移和TME的变化，如整体和肿瘤细胞特异性HGFL损失。我们还展示了一个 在人BC中协调HGFL和罗恩的上调。为了获得对罗恩信号传导的机制性见解， 为了研究RON通路，我们对罗恩调节的人和鼠BC细胞进行了RNA-Seq分析。令人惊讶的是，我们 发现罗恩主要抑制I型干扰素（IFN）、细胞因子和模式相关的途径。 识别受体（PRR）信号传导。通过研究这些途径，我们发现了一种新的联系， 罗恩和抑制IRAK 4（白细胞介素-1受体相关激酶4），一种已鉴定的先天性 PRR下游的免疫反应。我们还确定了IRF 7的减少，IRF 7是I型IFN的关键调节因子， 有罗恩丢失和IRAK 4表达。我们表明，IRAK 4导入表达罗恩的细胞中， 罗恩损失，而用IRAK 4抑制剂处理罗恩缺陷细胞， 在肿瘤生长和I型IFN产生方面。根据这些数据，本提案将检验假设 肿瘤细胞内源性罗恩的上调通过对肿瘤本身的作用和对肿瘤细胞的增殖的作用促进了BC的进展。 和TME通过涉及抑制IRAK 4和随后的I型IFN产生的机制。两个目标 （i）确定肿瘤细胞内在罗恩信号传导和I型IFN的功能意义 破坏建立肿瘤容许微环境以支持BC生长，以及（ii）定义 罗恩活化抑制IRAK 4-I型IFN信号传导轴以促进BC生长和转移。这 这项工作有望为攻击性BC的潜在机制提供新的见解，并将在以下方面具有指导意义： 获得联合疗法，指导临床研究，以防治BC死亡。

项目成果

Age-related changes in the epithelial and stromal compartments of the mammary gland in normocalcemic mice lacking the vitamin D3 receptor.

• DOI: 10.1371/journal.pone.0016479
• 发表时间: 2011-01-26
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Welsh J;Zinser LN;Mianecki-Morton L;Martin J;Waltz SE;James H;Zinser GM
• 通讯作者: Zinser GM

The human DEK oncogene stimulates β-catenin signaling, invasion and mammosphere formation in breast cancer.

• DOI: 10.1038/onc.2011.2
• 发表时间: 2011-06-16
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Vinnedge, L. M. Privette;McClaine, R.;Wagh, P. K.;Wikenheiser-Brokamp, K. A.;Waltz, S. E.;Wells, S. I.
• 通讯作者: Wells, S. I.

Ron receptor-dependent gene regulation in a mouse model of endotoxin-induced acute liver failure.

内毒素诱导的急性肝衰竭小鼠模型中 Ron 受体依赖性基因调控。

• DOI: 10.1016/s1499-3872(12)60196-9
• 发表时间: 2012
• 期刊: Hepatobiliary & pancreatic diseases international : HBPD INT
• 作者: Kulkarni,RishikeshM;Kutcher,LouisW;Stuart,WilliamD;Carson,DanielJ;Leonis,MikeA;Waltz,SusanE
• 通讯作者: Waltz,SusanE