CCAR2 as a Target for Prevention of Colorectal Cancer.

CCAR2 作为预防结直肠癌的靶点。

• 批准号: 10565953
• 负责人: Roderick H Dashwood
• 金额: $ 54.83万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10565953
• 关键词: Acetylation; Acute; Aftercare; Alternative Splicing; Antibodies; Apoptosis; Apoptosis Regulation Gene; Attenuated; Binding; Biopsy; Bromodomain; CRISPR/Cas technology; Cell Cycle; Cell Cycle Proteins; Cell Death; Cell Survival; Clinical; Colon; Colon Carcinoma; Colorectal Cancer; Complex; Cyclin D1; Dietary Factors; Dietary Isothiocyanate; Dose; Down-Regulation; Drug Combinations; Endoscopy; Engineering; Excision; Familial Adenomatous Polyposis Syndrome; Genetic Transcription; HDAC3 gene; Histone Deacetylase Inhibitor; Human; Individual; Induction of Apoptosis; Isothiocyanates; Link; Modeling; Nuclear; Nude Mice; Null Lymphocytes; Patients; Polyps; Prevention Protocols; Preventive; Prognosis; Protein Family; Proteins; RNA Binding; RNA Processing; RNA Splicing; Rattus; Reader; Role; Secondary Prevention; Signal Pathway; Signal Transduction; Site; Sulforaphane; Tertiary Protein Structure; Testing; Transfection; Transplantation; Tumor Suppression; c-myc Genes; cancer cell; colon cancer patients; colon cancer prevention; colorectal cancer prevention; cruciferous vegetable; dietary; experimental study; in vivo; inhibitor; member; mutant; novel; overexpression; patient derived xenograft model; polyposis; protein protein interaction; synergism; tumor

Cell Cycle and Apoptosis Regulator Protein 2 (CCAR2) is overexpressed in human colorectal cancer and is associated with poor prognosis, due in part to its role as a coactivator of -catenin-dependent transcription. The dietary preventive agent sulforaphane (SFN) targets CCAR2 interacting protein histone deacetylase 3 (HDAC3) for inhibition, resulting in acetylation of CCAR2, reduced CCAR2/-catenin nuclear interactions, and attenuated -catenin-dependent transcription. The novel CCAR2 acetylation sites align with a domain linked to S1 RNA binding, providing a new mechanistic link to alternative RNA splicing by SFN and other dietary isothio- cyanates (ITCs). Using novel protein domain arrays, bromodomain-containing proteins were identified that recognized acetylated forms of CCAR2. These acetyl “readers” also were inhibited by JQ1, which binds to bromodomain and extraterminal (BET) family proteins, and synergized with SFN to inhibit cell viability in colon cancer cells. CENTRAL HYPOTHESIS: In the prevention of colon cancer by dietary ITCs and mechanistically- prioritized drug combinations, HDAC3 inhibition leads to acetylation of CCAR2 and changes in protein-protein interactions that enhance apoptosis via the inhibition of -catenin-dependent transcription and via altered RNA splicing. Aim 1a: Test the hypothesis that by inhibiting HDAC3, ITCs alter the acetylation status of CCAR2 in colon cancer cells, with consequences for -catenin signaling and apoptosis induction. Aim 1b: Perform mechanistic studies on the acetyl readers of CCAR2, and test inhibitors of readers that synergize with SFN. Aim 1c: Examine the role of CCAR2 and the DBIRD complex in RNA splicing. Aim 2a: Using the polyposis in rat colon (Pirc) model, extend to working hypothesis in vivo linking HDAC3 inhibition, CCAR2 acetylation, reduced -catenin signaling and apoptosis induction. Single acute doses of SFN, 6-SFN and 9-SFN will be tested alone and in combination with JQ1. The most effective agents from Aim 2a will be assessed for tumor suppression in the Pirc model via primary (Aim 2b) and secondary prevention protocols (Aim 2c). Using sequential endoscopy and polyp resection in live rats, define the precise timing of HDAC3 protein loss, CCAR2 acetylation, reduced nuclear CCAR2/-catenin interactions, and the downregulation of -catenin targets. Aim 2d: Extend mechanistic studies from Aim 1c linking CCAR2 acetylation to alternative RNA splicing after ITC treatment into the Pirc model. Aim 3a: In biopsies from FAP patients, establish the translational relevance of findings in the Pirc model with respect to CCAR2 overexpression and its interactions with -catenin, HDAC3, and ZIRD. Aim 3b: In colonoids from Pirc and FAP patients, test the hypothesis that SFN, 6-SFN and 9-SFN alter the acetylation status of CCAR2 and its protein-protein interactions, with consequences for -catenin signaling and apoptosis induction. Test individual ITCs and their combinations with JQ1. Aim 3c: In nude mice transplanted with FAP patient-derived xenografts or CRISPR/Cas9-engineered CCAR2-null human colon cancer cells transfected with CCAR2 WT and acetylation mutants, examine tumor suppression by ITCs±JQ1.

细胞周期和细胞凋亡调节蛋白 2 (CCAR2) 在人类结直肠癌中过度表达，并且 与不良预后相关，部分原因是其作为 β-连环蛋白依赖性转录的共激活剂的作用。 饮食预防剂萝卜硫素 (SFN) 靶向 CCAR2 相互作用蛋白组蛋白脱乙酰酶 3 (HDAC3) 进行抑制，导致 CCAR2 乙酰化，减少 CCAR2/-连环蛋白核相互作用，以及 减弱的 -连环蛋白依赖性转录。新的 CCAR2 乙酰化位点与连接到的结构域对齐 S1 RNA 结合，为 SFN 和其他饮食异硫代RNA选择性剪接提供新的机制链接 氰酸盐 (ITC)。使用新型蛋白质结构域阵列，鉴定出含溴结构域的蛋白质 公认的 CCAR2 乙酰化形式。这些乙酰基“阅读器”也受到 JQ1 的抑制，JQ1 与 溴结构域和末端外 (BET) 家族蛋白，并与 SFN 协同抑制结肠细胞活力 癌细胞。中心假设：通过膳食 ITC 和机制来预防结肠癌—— 优先药物组合，HDAC3抑制导致CCAR2乙酰化和蛋白质-蛋白质变化 通过抑制 β-连环蛋白依赖性转录和改变 RNA 来增强细胞凋亡的相互作用 拼接。目标 1a：检验以下假设：ITC 通过抑制 HDAC3 改变 CCAR2 的乙酰化状态 结肠癌细胞，对 β-连环蛋白信号传导和细胞凋亡诱导产生影响。目标 1b：执行 对CCAR2的乙酰基阅读器进行机制研究，并测试与SFN协同的阅读器抑制剂。 目标 1c：检查 CCAR2 和 DBIRD 复合物在 RNA 剪接中的作用。目标 2a：利用息肉病 大鼠结肠 (Pirc) 模型，扩展到体内连接 HDAC3 抑制、CCAR2 乙酰化、 减少-连环蛋白信号传导和细胞凋亡诱导。单次急性剂量的 SFN、6-SFN 和 9-SFN 将 单独测试以及与 JQ1 结合测试。将评估 Aim 2a 中最有效的药物对肿瘤的影响 Pirc 模型中通过一级预防（目标 2b）和二级预防方案（目标 2c）进行抑制。使用 对活体大鼠进行连续内窥镜检查和息肉切除术，确定 HDAC3 蛋白丢失、CCAR2 的精确时间 乙酰化、核 CCAR2/-连环蛋白相互作用减少以及 -连环蛋白靶点下调。目的 2d：从 Aim 1c 延伸机制研究，将 CCAR2 乙酰化与 ITC 后的替代 RNA 剪接联系起来 处理入 Pirc 模型。目标 3a：在 FAP 患者的活检中，确定以下因素的转化相关性： Pirc 模型中有关 CCAR2 过度表达及其与 β-连环蛋白、HDAC3、 和ZIRD。目标 3b：在 Pirc 和 FAP 患者的结肠样中，检验 SFN、6-SFN 和 9-SFN 的假设 改变 CCAR2 的乙酰化状态及其蛋白质-蛋白质相互作用，对 -catenin 产生影响 信号传导和细胞凋亡诱导。使用 JQ1 测试单个 ITC 及其组合。目标 3c：在裸鼠中 移植 FAP 患者来源的异种移植物或 CRISPR/Cas9 工程改造的 CCAR2 缺失人类结肠 用 CCAR2 WT 和乙酰化突变体转染的癌细胞，检查 ITC±JQ1 的肿瘤抑制作用。

项目成果

CCAR1 and CCAR2 as gene chameleons with antagonistic duality: Preclinical, human translational, and mechanistic basis.

• DOI: 10.1111/cas.14579
• 发表时间: 2020-10
• 期刊: Cancer science
• 影响因子: 5.7
• 作者: Johnson GS;Rajendran P;Dashwood RH
• 通讯作者: Dashwood RH

Long noncoding RNAs and sulforaphane: a target for chemoprevention and suppression of prostate cancer.

• DOI: 10.1016/j.jnutbio.2017.01.001
• 发表时间: 2017-04
• 期刊: The Journal of nutritional biochemistry
• 作者: Beaver LM;Kuintzle R;Buchanan A;Wiley MW;Glasser ST;Wong CP;Johnson GS;Chang JH;Löhr CV;Williams DE;Dashwood RH;Hendrix DA;Ho E
• 通讯作者: Ho E

BRD9 Inhibition by Natural Polyphenols Targets DNA Damage/Repair and Apoptosis in Human Colon Cancer Cells.

• DOI: 10.3390/nu14204317
• 发表时间: 2022-10-15
• 期刊: NUTRIENTS
• 影响因子: 5.9
• 作者: Kapoor, Sabeeta;Damiani, Elisabetta;Wang, Shan;Dharmanand, Ravirajan;Tripathi, Chakrapani;Perez, Jorge Enrique Tovar;Dashwood, Wan Mohaiza;Rajendran, Praveen;Dashwood, Roderick Hugh
• 通讯作者: Dashwood, Roderick Hugh

NADPH oxidase overexpression in human colon cancers and rat colon tumors induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP).

• DOI: 10.1002/ijc.25610
• 发表时间: 2011-06-01
• 期刊: INTERNATIONAL JOURNAL OF CANCER
• 影响因子: 6.4
• 作者: Wang, Rong;Dashwood, Wan-Mohaiza;Nian, Hui;Loehr, Christiane V.;Fischer, Kay A.;Tsuchiya, Naoto;Nakagama, Hitoshi;Ashktorab, Hassan;Dashwood, Roderick H.
• 通讯作者: Dashwood, Roderick H.

Divergent roles of p120-catenin isoforms linked to altered cell viability, proliferation, and invasiveness in carcinogen-induced rat skin tumors.

• DOI: 10.1002/mc.22630
• 发表时间: 2017-07
• 期刊: Molecular carcinogenesis
• 影响因子: 4.6
• 作者: Wang R;Chen YS;Dashwood WM;Li Q;Löhr CV;Fischer K;Ho E;Williams DE;Dashwood RH
• 通讯作者: Dashwood RH