Mapping p53 dynamics to cell-fate outcomes in reprogramming and oncogenesis

将 p53 动态映射到重编程和肿瘤发生中的细胞命运结果

• 批准号: 10744532
• 负责人: Adam Matthew Beitz
• 金额: $ 4.92万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-11 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10744532
• 关键词: Address; Adopted; Attention; Back; Behavior; Binding; Bypass; Cancer Model; Cancerous; Cell Differentiation process; Cell Fraction; Cell Reprogramming; Cell Separation; Cells; DNA Binding Domain; Diagnosis; Disease; Early Diagnosis; Ectoderm; Endoderm; Engineering; Epigenetic Process; Epithelial Cells; Epithelium; Event; Fibroblasts; Frequencies; Generations; Genes; Genetic Transcription; Genomics; Human; Human Development; Immune; Investigation; Lead; Link; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Oviducts; Mammary Gland Parenchyma; Maps; Mediating; Mesoderm; Modeling; Molecular; Motor Neurons; Mus; Mutate; Mutation; Neoplasm Metastasis; Neurons; Oncogenes; Oncogenic; Organism; Organoids; Outcome; Patient-Focused Outcomes; Phase; Phenotype; Pluripotent Stem Cells; Point Mutation; Postdoctoral Fellow; Process; Proliferating; Proteins; Regulation; Reporter; Reporting; Research; Role; Sampling; Skin; Somatic Cell; Stains; Synthetic Genes; System; TP53 gene; Therapeutic; Tissues; Tumor Suppressor Proteins; Work; cancer initiation; carcinogenesis; cell type; design; effective therapy; enhancing factor; gain of function; improved; induced pluripotent stem cell; mutant; overexpression; prevent; programs; sensor; small hairpin RNA; stem cell differentiation; stem cell fate; stem cell fate specification; stem cells; success; targeted treatment; three-dimensional modeling; transcription factor; trend; tumor; tumor initiation; tumor progression; tumorigenesis

Project Summary Cell fates are decided as an organism develops. In human development, pluripotent stem cells differentiate into the three layers of ectoderm, mesoderm, and endoderm. These classes of tissue further differentiate into specific cell types with specific functions including neurons, immune cells, and skin cells. These identities are stable; once a cell differentiates into its final state, it will not revert back to a stem cell state, nor will it transform into another cell type. A skin cell will not spontaneously become a neuron, even if the neuron is damaged. However, Takahashi and Yamanaka demonstrated that cells have the potential to revert back to a stem cell fate when they reprogrammed mouse fibroblasts into induced pluripotent stem cells (iPSCs) by forced overexpression of stem cell-specifying transcription factors. In 2010, Vierbuchen and colleagues demonstrated that fibroblasts could be reprogrammed directly to neurons using neuron-specific transcription factors, bypassing the need for an iPSC-intermediate. However, reprogramming efficiencies in each of these systems was low; very few cells are actually capable of changing their cellular identity. In 2019, Babos and Galloway greatly improve reprogramming efficiencies in direct motor neuron reprogramming, demonstrating improved reprogramming yields 100 times greater than the original process. They drew upon factors that enhanced another cell fate transition: cancer. Genes that promote a healthy cell’s transition to cancer also improved the ability of a cell to change its cell type. Thus, reprogramming can serve as a model of cancer initiation. By understanding the molecular mechanisms by which these oncogenes promote reprogramming, we can understand how oncogenes evade cellular barriers to cancer and establish tumors. In the F99-phase of the proposed research, I will investigate the role of the tumor suppressor protein p53 in oncogene-mediated reprogramming. p53 is the most frequently mutated gene in cancer. Rather than p53 expression being lost in cancer, it is most often mutated to create a protein unable to perform its designated functions and accumulates to abnormally high levels. As a synthetic biologist, I will design synthetic gene circuits that track and report p53 levels during reprogramming. I will isolate cells that accumulate p53 and investigate their ability to reprogram. In the K00-phase of the proposed research, I will extend my investigations of p53 to three-dimensional models of ovarian cancer. Ovarian cancer is often diagnosed at late stages, after the cancer has metastasized, leading to poor patient outcomes. 3D models of tumor initiation can shed light on the early stages of ovarian cancer and enable clinicians to catch the cancer early, when the disease is most easily treated. By inducing cancer initiation in 3D models of ovarian cancer and tracking cancer progression using p53-sensors, I will identify the drivers of tumor establishment and factors associated with early-stage disease.

项目摘要 细胞命运被确定为生物体的发展。在人类发育中，多能干细胞 分化为外胚层，中胚层和内胚层的三层。这些组织进一步 通过特定功能区分特定的细胞类型，包括神经元，免疫细胞和皮肤细胞。这些 身份稳定；一旦细胞区分到最终状态，它将不会恢复到干细胞状态，也不会 它转变为另一种单元格类型。即使神经元是，皮肤细胞也不会成为神经元 损坏的。但是，高桥和山内卡证明了细胞有可能恢复到 干细胞的命运将小鼠成纤维细胞重编程为诱导多能干细胞（IPSC）时 干细胞特异性转录因子的过表达。 2010年，Vierbuchen及其同事证明了 可以使用神经特异性转录因子将成纤维细胞直接重编程为神经元 需要IPSC中级。但是，每个系统中的每个系统的重编程效率都很低。非常 实际上，很少有细胞能够改变其细胞身份。 在2019年，Babos和Galloway极大地提高了直接运动神经元的重编程效率 重新编程，证明重新编程的改进，比原始过程大100倍。 他们借鉴了增强了另一种细胞命运转变的因素：癌症。促进健康细胞的基因 过渡到癌症还提高了细胞改变其细胞类型的能力。那就是重新编程可以作为 癌症开始的模型。通过了解这些癌基因促进的分子机制 重新编程，我们可以理解Oncogenes如何逃避癌症的细胞障碍并建立肿瘤。 在拟议的研究的F99期间，我将研究肿瘤抑制蛋白p53在 癌基因介导的重编程。 p53是癌症中最常见的突变基因。而不是p53 表达在癌症中流失，通常会突变以创建无法执行其指定的蛋白质 功能并累积到绝对高水平。作为合成生物学家，我将设计合成基因电路 在重新编程过程中，该跟踪和报告P53水平。我将分离积累p53并研究的细胞 他们重新编程的能力。 在拟议的研究的K00期间，我将把p53的研究扩展到三维 卵巢癌的模型。癌症转移后，经常在晚期诊断出卵巢癌， 导致患者的结果差。肿瘤起始的3D模型可以揭示卵巢的早期阶段 当疾病最容易治疗时，癌症并使临床医生能够尽早感染癌症。由诱导 在卵巢癌的3D模型中的癌症开始，并使用p53传感器跟踪癌症的进展，我将确定 肿瘤的驱动因素和与早期疾病相关的因素。