SPROUTY: PUTATIVE ONCOGENE IN COLORECTAL CANCER

结直肠癌（CRC）是美国癌症相关死亡的主要原因。 结直肠癌的死亡主要是由于肿瘤复发和远处转移。Sprouty2 （SPRY 2），受体酪氨酸激酶（RTK）/促分裂原活化的内源性抑制剂 蛋白激酶（MAPK）信号通路，作为乳腺癌，前列腺癌， 肝癌我们发现SPRY 2在CRC中的致癌作用。本申请的目的 评估SPRY 2在CRC进展、治疗和预后中的需要和意义。 复发 研究表明，SPRY 2的上调加重了癌症表型， SPRY 2的抑制增强了上皮特征并减少了上皮间充质 过渡（EMT）。我们假设SPRY 2的缺失将抑制CRC。为了验证这一 假设，在AIM 1中，将利用SPRY 2 LoxP/Vil-Cre ERT 2小鼠模型。SPRY 2的影响 将研究缺失对氧化偶氮甲烷（AOM）诱导的结肠肿瘤发生的影响。据进一步 证实SPRY 2稳定转染显著增加RTK cMet表达。 抑制SPRY 2降低内源性和TGF-β诱导的cMet蛋白和mRNA 表情在AIM 2中，我们将描述TGF-β 1/SPRY 2依赖的转录调控， 结肠癌细胞系中的cMET。虽然表皮生长因子受体（EGFR）的抑制剂是 在转移性CRC的治疗中，cMet表达和EMT的增加是有益的。 抗EGFR治疗的耐药机制。在AIM 3中，SPRY 2抑制对肿瘤的影响 吉非替尼和西妥昔单抗的生长（异种移植模型）和转移（盲肠植入模型）- 将研究抗性结肠癌细胞系。我们假设SPRY 2抑制将 使吉非替尼和西妥昔单抗抗性结肠癌细胞对抗EGFR疗法敏感。研究将 扩展到分析SPRY 2在体内吉非替尼治疗过程中缺失的影响 LoxP/Vil-Cre ERT 2小鼠模型。SPRY 2在肿瘤复发及远期预后中的意义 生存并不明确。在AIM 4中，我们假设SPRY 2在原发性肿瘤中的表达可能 与肿瘤复发和长期生存直接相关。此外，由于SPRY 2是 Ras依赖性途径的调节剂，我们已经扩展了我们的调查，以评估 SPRY 2表达与Ras突变[KRAS（密码子12，13）和NRAS（密码子14，15）]的关联 12，13）]在复发。SPRY 2表达和Ras突变的共同存在在乳腺癌中的意义 将研究原发性肿瘤与疾病结局的关系。对于该分析，散发性II-III期 存档的福尔马林固定石蜡包埋肿瘤（FFPET），与5年时的结局相关 将被利用。 据我们所知，这是第一项证明SPRY 2在癌症中的促癌作用的研究。 《儿童权利公约》。我们根据初步数据和出版物建立了假设。的 提出的研究是一种多管齐下的方法来评估SPRY 2缺失对肿瘤的影响， 生长，检查SPRY 2在获得性抗EGFR治疗耐药性中的作用，并评估SPRY 2在获得性抗EGFR治疗耐药性中的作用。 SPRY 2和Ras突变在鉴别肿瘤风险增加患者中的意义 复发

Colorectal cancer (CRC) is the leading cause of cancer-related death in the United States. Cancer death in CRC is mainly due to tumor recurrence and distant metastasis. Sprouty2 (SPRY2), an endogenous suppressor of receptor tyrosine kinase (RTK)/ Mitogen Activated Protein Kinase (MAPK) signaling pathways, acts as a tumor suppressor in breast, prostate, and liver cancer. We discovered oncogenic role of SPRY2 in CRC. The objectives of this application are to assess the requirement and significance of SPRY2 in CRC progression, treatment and recurrence. Studies demonstrated that upregulation of SPRY2 accentuates cancer phenotype whereas suppression of SPRY2 augments epithelial features and decreases epithelial mesenchymal transition (EMT). We hypothesize that deletion of SPRY2 will suppress CRC. To test this hypothesis, in AIM 1, SPRY2 LoxP/Vil-Cre ERT2 mouse model will be utilized. Effect of SPRY2 deletion on azoxymethane (AOM)-induced colonic tumorigenesis will be studied. It was further established that SPRY2 stable transfection significantly increased RTK cMet expression. Suppression of SPRY2 decreased endogenous and TGF- induced cMet protein and mRNA expression. In AIM 2, we will delineate TGF-/SPRY2 dependent transcriptional regulation of cMET in colon cancer cell lines. While inhibitors of epidermal growth factor receptor (EGFR) are beneficial in the treatment of metastatic CRC, increases in cMet expression and EMT are resistance mechanism to anti-EGFR therapy. In AIM 3, effect of SPRY2 suppression on tumor growth (xenograft model) and metastasis (cecal implantation model) of gefitinib and cetuximab- resistant colon cancer cell lines will be investigated. We hypothesize that SPRY2 suppression will sensitize gefitinib and cetuximab-resistant colon cancer cells to anti-EGFR therapy. Studies will be extended to analyze effect of SPRY2 deletion during gefitinib treatment in vivo SPRY2 LoxP/Vil-Cre ERT2 mouse model. Significance of SPRY2 with tumor recurrence and long-term survival is not clear. In AIM 4, we hypothesize that SPRY2 expression in primary tumors may correlate directly with tumor recurrence and long-term survival. Further, as SPRY2 is the key regulator of Ras-dependent pathways, we have extended our investigations to assess the association of SPRY2 expression with Ras mutations [KRAS (codon 12, 13) and NRAS (codon 12, 13)] in recurrence. Implication of concomitance of SPRY2 expression and Ras mutations in primary tumors to disease outcome will be investigated. For this analysis sporadic stage II-III archived formalin-fixed paraffin embedded tumors (FFPET) that are linked to outcomes at 5 years will be utilized. To our knowledge, this is the first study to demonstrate a cancer promoting role of SPRY2 in CRC. We have established hypotheses based on our preliminary data and publications. The proposed study is a multi-pronged approach to assess the effect of SPRY2 deletion on tumor growth, examine the role of SPRY2 in acquired resistance to anti-EGFR therapy and evaluate the significance of SPRY2 and Ras mutations in identifying patients with increased risk of tumor recurrence.