p53, Aging, and Cancer

p53，衰老与癌症

• 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
• 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' 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

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.

在已建立的CD19+白血病模型中，已经检查了具有Delta133P53Alpha表达盒的CAR-T细胞的抗肿瘤活性。与对照CAR-T细胞相比，在体外和体内的小鼠注射实验中，delta133p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53p53揭示了这种优质活性与p53介导的衰老和凋亡的抑制（维持p53介导的DNA修复）以及代谢转移，例如线粒体氧化磷酸化和核糖体生物发生等代谢转移，这两者都有利于T细胞干，增殖和持续活性。重要的是，我们观察到Delta133p53Alpha诱导的功能性改善不仅是从正常供体衍生的CAR-T细胞中，而且还来自CLL（慢性淋巴细胞性白血病）患者的患者，这些患者对当前CAR-T细胞治疗没有反应，从而支持其临床治疗价值。这些数据需要进一步研究这种新型的CAR-T细胞策略，以治疗当前的非反应者和目前难以治疗的实体瘤患者。为了研究delta133p53alpha的体内治疗作用，我们生成了多个表达人/灵长类特异性的delta133p53alpha同工型的小鼠模型。由于人类p53敲入小鼠（hupki）在人类p53敲击的小鼠（hupki）中的谦虚表达对后代模型小鼠的加速衰老表型没有任何影响，因此我们生成了避风势 - 玛尔的转基因小鼠，在其中delta133p53alpha中，在同步的rouseratient of the anttecters oferant的启动器（强度）中，或通过CRE/ERT2-LOXP系统诱导。在他莫昔芬治疗后，这些转基因小鼠在各种器官中表达delta133p53alpha。他们目前正在衰老研究中，以检查生理衰老过程和自发性肿瘤的发病率和频谱，并且还与早期模型小鼠一起繁殖，以检查对加速衰老表型的影响。这些小鼠也是我们对神经退行性疾病的研究的重要材料，它通过阿尔茨海默氏病模型小鼠繁殖并采用诸如颅骨辐射（模拟放射治疗引起的晚期认知疾病的癌症幸存者中的晚期认知障碍）等方案（模仿慢性卫生型运动员）和慢性运动型运动员。我们已经对小分子文库，重新利用的药物和天然产物进行了高通量筛选，以识别可以增强Delta133p53alpha表达的候选药物，从而导致两个候选者（化合物“ A'''和化合物” C'，在此无法在此处透露身份）。他们被证实会增加Delta133p53Alpha的表达，并抑制与衰老的衰老星形胶质细胞和衍生的成纤维细胞中与衰老相关的分泌表型。 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.由于这些化合物已经用于安全性和其他疾病的临床试验中，因此它们可以显着促进我们将实验室发现转化为诊所。我们通过研究p53同工型p53beta和delta133p53alpha的癌症相关突变版，开创了p53研究的新领域。虽然先前证明了剪接因子SRSF3的siRNA敲低可以诱导p53beta（通过替代剪接从全长p53转换为p53转换为p53beta）和p53野生型正常细胞中的细胞衰老，我们最近发现，在p53-Mutant gbm和lung conget p53中，我们也发现了p53-含量AnockES p5 connect p p.3 opta p p.3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的全长P53转换为肿瘤抑制突变突变体P53Beta同工型，促使我们开发了一种针对癌症特异性衰老和无针对正常细胞的癌症特异性衰老和极端影响的SRSF3 siRNA的癌症特异性递送方法。我们还揭示了野生型和突变体delta133p53p53Alpha同工型在GBM细胞中功能截然不同。突变的Delta133p53Alpha通过增加细胞增殖和侵袭，DNA修复受损和激活的IL4I1/IDO1/AHR途径来致癌，这可以在p53-突动剂GBM中成为治疗靶标。 [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. CellRep。42：112260，2023。

项目成果

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

• DOI: 10.1038/onc.2012.288
• 发表时间: 2013-05-30
• 期刊: Oncogene
• 影响因子: 8
• 作者: Tang Y;Horikawa I;Ajiro M;Robles AI;Fujita K;Mondal AM;Stauffer JK;Zheng ZM;Harris CC
• 通讯作者: Harris CC

EML4-ALK induces cellular senescence in mortal normal human cells and promotes anchorage-independent growth in hTERT-transduced normal human cells.

• DOI: 10.1186/s12885-021-07905-6
• 发表时间: 2021-03-24
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: Miyanaga A;Matsumoto M;Beck JA;Horikawa I;Oike T;Okayama H;Tanaka H;Burkett SS;Robles AI;Khan M;Lissa D;Seike M;Gemma A;Mano H;Harris CC
• 通讯作者: Harris CC

Astrocyte senescence and SASP in neurodegeneration: tau joins the loop.

神经变性中的星形胶质细胞衰老和 SASP：tau 蛋白加入环路。

• DOI: 10.1080/15384101.2021.1909260
• 发表时间: 2021
• 期刊: Cell cycle (Georgetown, Tex.)
• 作者: Ungerleider,Kyra;Beck,Jessica;Lissa,Delphine;Turnquist,Casmir;Horikawa,Izumi;Harris,BrentT;Harris,CurtisC
• 通讯作者: Harris,CurtisC

Telomerase variant A279T induces telomere dysfunction and inhibits non-canonical telomerase activity in esophageal carcinomas.

端粒酶变体 A279T 会诱导端粒功能障碍并抑制食管癌中的非典型端粒酶活性。

• DOI: 10.1371/journal.pone.0101010
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Zhang,Yuwei;Calado,Rodrigo;Rao,Mahadev;Hong,JulieA;Meeker,AlanK;Dumitriu,Bogdan;Atay,Scott;McCormick,PeterJ;Garfield,SusanH;Wangsa,Danny;Padilla-Nash,HesedM;Burkett,Sandra;Zhang,Mary;Kunst,TriciaF;Peterson,NathanR;Xi,
• 通讯作者: Xi,

Corrigendum to: The role of p53 in base excision repair following genotoxic stress.

勘误表：p53 在基因毒性应激后碱基切除修复中的作用。

• DOI: 10.1093/carcin/bgab028
• 发表时间: 2021
• 期刊: Carcinogenesis
• 影响因子: 4.7
• 作者: Zurer,Irit;Hofseth,LorneJ;Cohen,Yehudit;Xu-Welliver,Meng;Hussain,SPerwez;Harris,CurtisC;Rotter,Varda
• 通讯作者: Rotter,Varda