Functions of Cyclin/CDK complexes in development and cancer

项目摘要 彼得·西辛斯基博士在达纳-法伯癌症研究所的实验室研究了细胞周期蛋白在 在正常发育和癌症中。在过去的20年里，我们创造了许多鼠标击倒或敲入 菌株包括缺乏周期蛋白或CDK的小鼠，或表达修饰的周期蛋白或CDK的小鼠。使用那些 小鼠，我们描绘了正常小鼠发育和肿瘤中对细胞周期蛋白的需求 形成/发展。作为一名研究和分析转基因小鼠模型的专家。 研究专家严庚一直在领导西辛斯基博士的实验室努力创造不同的 Cyclin/CDK突变株。尤其是，她在以下领域发挥了关键作用：1)设计 针对Cyclin基因敲除或Cyclin或CDK基因敲入突变/标签的基因靶向策略 2)监督胚胎干细胞基因操作的技术过程和创造 胚胎干细胞植入嵌合体小鼠；3)利用小鼠肿瘤设计并进行实验 模型包括异种/同种移植模型。目前，研究专家正在制造一种新的老鼠品系 条件基因敲除CDK19。利用这种菌株，我们计划研究cdk19缺失的影响与 T细胞白血病中细胞周期蛋白C的缺失。结合CDK19基因敲除和细胞周期蛋白C基因敲除，我们希望创造一种 忠实模仿人类T细胞白血病的小鼠模型。研究专家也在领导 G1期细胞周期蛋白在癌细胞干细胞分化中的作用研究我们已经发现，G1期细胞周期蛋白直接调节 多能因子Oct4、Sox2和Nanog的蛋白水平不仅在胚胎干细胞中，而且在 胶质母细胞瘤肿瘤起始细胞，我们将研究是否降低G1周期蛋白的水平/活性 会对肿瘤细胞的干性产生影响。我们将使用异种移植模型来测试击倒 G1期细胞周期蛋白将诱导胶质母细胞瘤细胞分化，从而消除/降低其形成的潜能 体内的肿瘤。最近，我们发现细胞周期蛋白D/CDK4/6在肿瘤免疫中发挥作用。使用肿瘤 同种异体小鼠移植模型，研究专家将探索是否抑制细胞周期蛋白D/CDK4/6 会增强抗免疫检查点阻断治疗黑色素瘤的疗效。我们希望这一点 这项研究将为黑色素瘤患者带来一种新的联合治疗策略。

Project Summary Dr. Peter Sicinski's laboratory at the Dana-Farber Cancer Institute studies the functions of cell cycle proteins in normal development and in cancer. For the past 20 years, we have created many mouse knockout or knock-in strains including mice either lacking cyclins or CDKs, or expressing modified cyclins or CDKs. Using those mice, we delineated the requirement for cell cycle proteins during normal mouse development and in tumor formation/progression. As an expert in generating and analyzing genetically engineered mouse models, Dr. Yan Geng, the Research Specialist, has been leading the efforts in Dr. Sicinski's lab to create different cyclin/CDK mutant strains. Particularly, she has played a crucial role in the following areas: 1) design the genetic targeting strategies for knockout cyclin genes or knock-in mutations/tags into a cyclin or CDK gene locus; 2) supervise the technical processes for gene manipulation in embryonic stem (ES) cells and creating chimeric mice by blastocyst injection of ES cells; 3) design and perform the experiments using mouse tumor models including xenograft/allograft models. Currently, the Research Specialist is making a new mouse strain of conditional knockout Cdk19. Using this strain, we plan to investigate the effect of Cdk19 loss together with loss of cyclin C in T-cell leukemia. Combining Cdk19 knockout with cyclin C knockout, we hope to create a mouse model that faithfully mimicking human T-cell leukemia. The Research Specialist is also leading the study of G1 cyclins' function in cancer cell stemness. As we have found that G1 cyclins directly regulate the protein levels of pluripotency factors Oct4, Sox2 and Nanog not only in embryonic stem cells but also in glioblastoma tumor initiating cells, we will investigate whether decreasing the levels/activities of G1 cyclins would impact on the stemness of tumor cells. We will use a xenograft model to test whether knocking down the G1 cyclins will induce differentiation of glioblastoma cells thus to eliminate/reduce their potential to form tumors in vivo. Recently, we have found that cyclin D/CDK4/6 play roles in cancer immunity. Using tumor allograft mouse models, the Research Specialist is going to explore whether inhibition of cyclin D/CDK4/6 would enhance the efficacy of anti-immune checkpoint blockade in treatment of melanoma. We hope that this study will lead to a new combinational therapeutic strategy for melanoma patients.