p53, Aging, and Cancer

p53，衰老与癌症

基本信息

批准号：
10926230
负责人：
Curtis Harris
金额：
$ 171.34万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
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/10926230
关键词：
Acceleration Adverse effects Aging Alternative Splicing Alzheimer&apos s disease model Apoptosis Area Astrocytes Biogenesis Biological Assay Brain Breeding CAR T cell therapy CD19 gene CD8-Positive T-Lymphocytes Cancer Survivor Cell Aging Cell Proliferation Cells Chronic Lymphocytic Leukemia Clinic Clinical Clinical Trials Coculture Techniques Cranial Irradiation Craniocerebral Trauma DNA Repair DNA cassette Data Development Disease Drug Screening Fibroblasts Generations Glioblastoma Human Impaired cognition Impairment In Vitro Incidence Injections Invaded Investigation Knock-in Mouse Laboratory Finding Length Malignant Neoplasms Malignant neoplasm of lung Mediating Metabolic Methods Mitochondria Mitotic spindle Modeling Mus Mutation Natural Products Nature Neurodegenerative Disorders Normal Cell Oncogenic Organ Oxidative Phosphorylation Pathology Pathway interactions Patients Phenotype Physiological Prevention Primates Process Progeria Proliferating Protein Isoforms Protocols documentation Publications RNA Splicing Radiation therapy Regulation Research Ribosomes Safety Small Interfering RNA Solid Neoplasm Syndrome System T-Lymphocyte TP53 gene Tamoxifen Therapeutic Therapeutic Effect Transgenic Mice Translations Tumor Suppression cancer cell cancer immunotherapy cancer type cell type chimeric antigen receptor T cells chronic traumatic encephalopathy clinical application contact sports drug candidate drug repurposing experimental study functional improvement high throughput screening humanized mouse improved in vivo in vivo Model knock-down leukemia lung cancer cell military veteran mouse model mutant non-oncogenic novel promoter senescence small molecule libraries stemness therapeutic target tumor

项目摘要

The CAR-T cells with the delta133p53alpha expression cassette have been examined for anti-tumor activity in an established CD19+ leukemia model. The delta133p53alpha-CAR-T cells showed superior tumor killing activity, compared with the control CAR-T cells, both in a co-culture experiment in vitro and in a mouse injection experiment in vivo. This superior activity was revealed to be associated with inhibition of p53-mediated senescence and apoptosis (while p53-mediated DNA repair was maintained) and metabolic shifts such as increased mitochondrial oxidative phosphorylation and ribosome biogenesis, which both favor T cell stemness, proliferation, and sustained activity. Importantly, we observed the delta133p53alpha-induced functional improvement not only in CAR-T cells derived from normal donors but also in those derived from CLL (chronic lymphocytic leukemia) patients who did not respond to the current CAR-T cell therapy, supporting its clinical therapeutic value. These data warrant further investigation of this novel CAR-T cell strategy toward treatment of current non-responders and patients with currently hard-to-treat solid tumors. To investigate in vivo therapeutic effects of delta133p53alpha, we have generated multiple mouse models that express the human/primate-specific delta133p53alpha isoform. Since modest expression of delta133p53alpha in human p53-knocked-in mice (hupki) did not show any effects on accelerated aging phenotypes in progeria model mice, we generated the safe-harbor transgenic mice in which delta133p53alpha under the control of the synthetic CAG promoter (strong) or the endogenous ROSA26 promoter (intermediate) can be induced via a Cre/ERT2-loxP system. These transgenic mice were confirmed to express delta133p53alpha in various organs upon tamoxifen treatment. They are currently under the aging study to examine physiological aging processes and spontaneous tumor incidence and spectrum, and also being bred with the progeria model mice to examine an effect on accelerated aging phenotypes. These mice are also an essential material for our study on neurodegenerative diseases, through breeding with Alzheimer's disease model mice and applying disease-causing protocols such as cranial irradiation (mimicking radiotherapy-induced late cognitive impairment in cancer survivors) and head trauma (mimicking chronic traumatic encephalopathy in contact sports athletes and military veterans). We have performed high-throughput screening of small molecule libraries, repurposed drugs and natural products to identify drug candidates that can enhance the expression of delta133p53alpha, leading to the two candidates (compound 'A' and compound 'C', whose identities cannot be disclosed here). They were confirmed to increase the delta133p53alpha expression and inhibit senescence-associated secretory phenotype in otherwise senescent astrocytes and progeria-derived fibroblasts. Other human cell types we previously used (e.g., CAR-T cells and endogenous CD8+ T cells) and the delta133p53alpha-humanized mouse models, as mentioned above, are being treated with these compounds to further confirm their activity and therapeutic potential. Since these compounds have already been used in clinical trials for safety and other diseases, they could significantly facilitate the translation of our laboratory findings to the clinic. We have pioneered a new area of p53 research by investigating cancer-associated mutant versions of the p53 isoforms p53beta and delta133p53alpha. While siRNA knockdown of a splicing factor SRSF3 was previously shown to induce p53beta (via alternative splicing switching from full-length p53 to p53beta) and cellular senescence in p53-wild-type normal human cells, we have recently discovered that, also in p53-mutant GBM and lung cancer cells, SRSF3 knockdown induces p53beta (with a cancer-associated mutation) and causes cellular senescence and apoptosis. These data suggest that SRSF3 knockdown can convert the oncogenic mutant full-length p53 to the tumor-suppressive mutant p53beta isoform, prompting us to develop a method of cancer-specific delivery of SRSF3 siRNA that aims at cancer-specific senescence and apoptosis with no or minimal adverse effect on normal cells. We have also revealed that wild-type and mutant delta133p53alpha isoforms are functionally distinct in GBM cells. Mutant delta133p53alpha functions oncogenic through increased cell proliferation and invasion, impaired DNA repair, and activated IL4I1/IDO1/AHR pathway, which can be a therapeutic target in p53-mutant GBM. [Publications] Muys BR, Shrestha RL, Anastasakis DG, Pongor L, Li XL, Grammatikakis I, Polash A, Chari R, Gorospe M, Harris CC, Aladjem MI, Basrai MA, Hafner M, Lal A. Matrin3 regulates mitotic spindle dynamics by controlling alternative splicing of CDC14B. Cell Rep. 42: 112260, 2023.

已在已建立的 CD19+ 白血病模型中检查了带有 delta133p53alpha 表达盒的 CAR-T 细胞的抗肿瘤活性。在体外共培养实验和体内小鼠注射实验中，与对照 CAR-T 细胞相比，delta133p53alpha-CAR-T 细胞显示出优异的肿瘤杀伤活性。研究表明，这种卓越的活性与抑制 p53 介导的衰老和细胞凋亡（同时维持 p53 介导的 DNA 修复）和代谢变化（例如增加线粒体氧化磷酸化和核糖体生物合成）有关，这两者都有利于 T 细胞干细胞性、增殖和增殖。持续的活动。重要的是，我们不仅在来自正常供体的 CAR-T 细胞中观察到了 delta133p53alpha 诱导的功能改善，而且在来自对当前 CAR-T 细胞疗法没有反应的 CLL（慢性淋巴细胞白血病）患者的细胞中也观察到了 delta133p53alpha 诱导的功能改善，支持其临床治疗价值。这些数据值得进一步研究这种新型 CAR-T 细胞策略，用于治疗当前无反应者和目前难以治疗的实体瘤患者。为了研究 delta133p53alpha 的体内治疗效果，我们构建了多个表达人类/灵长类动物特异性 delta133p53alpha 亚型的小鼠模型。由于 delta133p53alpha 在人 p53 敲入小鼠 (hupki) 中的适度表达并未显示出对早衰模型小鼠的加速衰老表型有任何影响，因此我们生成了安全港转基因小鼠，其中 delta133p53alpha 在合成 CAG 启动子的控制下（强）或内源性 ROSA26 启动子（中）可以通过 Cre/ERT2-loxP 系统诱导。这些转基因小鼠经他莫昔芬治疗后被证实在各个器官中表达 delta133p53alpha。它们目前正在进行衰老研究，以检查生理衰老过程和自发肿瘤的发病率和谱，并与早衰模型小鼠一起饲养，以检查对加速衰老表型的影响。这些小鼠也是我们研究神经退行性疾病的重要材料，通过与阿尔茨海默氏病模型小鼠进行繁殖并应用诸如颅脑照射（模仿放射治疗引起的癌症幸存者的晚期认知障碍）和头部创伤（模仿慢性创伤性创伤）等致病方案接触性运动运动员和退伍军人的脑病）。我们对小分子库、再利用药物和天然产物进行了高通量筛选，以确定可以增强 delta133p53alpha 表达的候选药物，从而产生了两种候选药物（化合物“A”和化合物“C”，其身份无法公开）这里）。它们被证实可以增加 delta133p53alpha 的表达，并抑制衰老星形胶质细胞和早衰源性成纤维细胞中与衰老相关的分泌表型。我们之前使用的其他人类细胞类型（例如 CAR-T 细胞和内源性 CD8+ T 细胞）和 delta133p53alpha 人源化小鼠模型（如上所述）正在用这些化合物进行处理，以进一步确认其活性和治疗潜力。由于这些化合物已用于安全性和其他疾病的临床试验，因此它们可以极大地促进我们的实验室研究结果转化为临床。我们通过研究 p53 同工型 p53beta 和 delta133p53alpha 的癌症相关突变版本，开创了 p53 研究的新领域。虽然剪接因子 SRSF3 的 siRNA 敲低先前被证明可诱导 p53beta（通过从全长 p53 到 p53beta 的选择性剪接转换）和 p53 野生型正常人类细胞中的细胞衰老，但我们最近发现，也在 p53-在突变 GBM 和肺癌细胞中，SRSF3 敲低会诱导 p53beta（具有与癌症相关的突变）并导致细胞衰老和凋亡。这些数据表明，SRSF3 敲低可以将致癌突变体全长 p53 转化为肿瘤抑制突变体 p53beta 同工型，促使我们开发一种癌症特异性递送 SRSF3 siRNA 的方法，该方法旨在实现癌症特异性衰老和细胞凋亡，而无需或对正常细胞的不良影响最小。我们还发现野生型和突变型 delta133p53alpha 亚型在 GBM 细胞中功能不同。突变体 delta133p53alpha 通过增加细胞增殖和侵袭、DNA 修复受损以及激活 IL4I1/IDO1/AHR 通路发挥致癌作用，这可以成为 p53 突变 GBM 的治疗靶点。 [出版物] Muys BR、Shrestha RL、Anastasakis DG、Pongor L、Li XL、Grammatikakis I、Polash A、Chari R、Gorospe M、Harris CC、Aladjem MI、Basrai MA、Hafner M、Lal A。Matrin3 调节有丝分裂纺锤体动力学通过控制 CDC14B 的选择性剪接。细胞报告 42：112260，2023。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Δ133p53α, a natural p53 isoform, contributes to conditional reprogramming and long-term proliferation of primary epithelial cells.

α133p53α 是一种天然的 p53 亚型，有助于原代上皮细胞的条件重编程和长期增殖。

DOI：
发表时间：
2018-07-03
期刊：
影响因子：
9
作者：
Mondal, Abdul M;Zhou, Hua;Horikawa, Izumi;Suprynowicz, Frank A;Li, Guangzhao;Dakic, Aleksandra;Rosenthal, Bernard;Ye, Lin;Harris, Curtis C;Schlegel, Richard;Liu, Xuefeng
通讯作者：
Liu, Xuefeng

Δ133p53 represses p53-inducible senescence genes and enhances the generation of human induced pluripotent stem cells.

α133p53 抑制 p53 诱导的衰老基因并增强人类诱导多能干细胞的生成。

DOI：
发表时间：
2017-06
期刊：
影响因子：
12.4
作者：
Horikawa, Izumi;Park, Kye;Isogaya, Kazunobu;Hiyoshi, Yukiharu;Li, Han;Anami, Katsuhiro;Robles, Ana I;Mondal, Abdul M;Fujita, Kaori;Serrano, Manuel;Harris, Curtis C
通讯作者：
Harris, Curtis C

Mutant p53 gain of function underlies high expression levels of colorectal cancer stem cells markers.

突变的 p53 功能获得是结直肠癌干细胞标记物高表达水平的基础。

DOI：
发表时间：
2018-03
期刊：
影响因子：
8
作者：
Solomon, Hilla;Dinowitz, Nathan;Pateras, Ioannis S;Cooks, Tomer;Shetzer, Yoav;Molchadsky, Alina;Charni, Meital;Rabani, Stav;Koifman, Gabriela;Tarcic, Ohad;Porat, Ziv;Kogan;Goldfinger, Naomi;Oren, Moshe;Harris, Curtis C;Gorgoulis
通讯作者：
Gorgoulis

Downregulation of splicing factor SRSF3 induces p53β, an alternatively spliced isoform of p53 that promotes cellular senescence.

剪接因子 SRSF3 的下调会诱导 p53β，这是一种促进细胞衰老的 p53 的选择性剪接亚型。