Defective PKA Signaling in Cushing's Syndrome

库欣综合征中 PKA 信号传导缺陷

• 批准号: 10582988
• 负责人: John D Scott
• 金额: $ 49.96万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-24 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10582988
• 关键词: A kinase anchoring protein; Adrenal Cortex; Adrenal Gland Neoplasms; Adrenal Glands; Alleles; Amendment; Anabolism; Arginine; Back; Biochemical; Blood Circulation; Blood Glucose; Catalytic Domain; Cell Nucleus; Cells; Chronic; Clinical; Cushing Syndrome; Cushingoid facies; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Defect; Development; Diabetes Mellitus; Disease; Endocrine; Endocrine System Diseases; Enzyme Stability; Enzymes; Event; Exclusion; Exposure to; Female; Glucose; Holoenzymes; Hormones; Hydrocortisone; Hypertension; Island; Knock-in Mouse; Left; Lesion; Molecular; Mus; Mutation; Operative Surgical Procedures; Organ Size; Other Genetics; Outcome; Pathogenesis; Pathogenicity; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Pituitary Neoplasms; Point Mutation; Proteins; Public Health; Radiation; Rosaniline Dyes; Sampling; Signal Pathway; Signal Transduction; Signaling Protein; Site; Stains; Stress; Symptoms; Testing; Tissues; Variant; Weight Gain; Woman; adenoma; adrenal hyperplasia; blood pressure elevation; chemotherapy; comorbidity; disease-causing mutation; intracellular protein transport; men; mouse model; mutant; precision medicine; protein distribution; restoration; spatiotemporal; transmission process; virtual

ABSTRACT Cushing’s syndrome occurs when the adrenal glands release too much of the stress hormone cortisol. This endocrine disorder is naturally caused by adrenal or pituitary tumors and afflicts women five times more frequently than men. Chronic exposure to cortisol also increases blood glucose and elevates blood pressure. This proposal investigates the molecular pathogenesis of adrenal Cushing’s syndrome. This disease is often an outcome of defective cAMP signaling in the adrenal cortex. Advances in precision medicine have identified point mutations and insertions in protein kinase A catalytic subunits (PKAc) as drivers of this disease. The most prevalent mutant is PKAc-L205R, which is found in ~45% of Cushing’s patients. Other genetic lesions include PKAc-W196R. Both disease-causing mutations reside within a region of the kinase domain that interfaces with its regulatory subunits. This protein-protein interface is necessary for autoinhibition of kinase activity, but also directs the subcellular targeting of PKA holoenzymes through association with A kinase anchoring proteins (AKAPs). Our biochemical, cell-based, and clinical analyses infer that carrying a mutant allele of PKAc-L205R or W196R is sufficient to displace the kinase from normal subcellular sites of action in Cushing’s patients. This has led to a working hypothesis that losing spatiotemporal control of protein kinase A underlies adrenal Cushing’s syndrome. Aim 1 will test if i) restoration of PKA anchoring amends cortisol release, ii) identify which AKAPs coordinate cortisol biosynthesis and iii) considers an opposing premise that “substrate rewiring” of PKAc variants or altered enzyme stability drives Cushing’s syndrome. At first glance, the PKAc-L205R and PKAc-W196R mutants look virtually identical. Yet we have discovered that these are physiochemically distinct kinases that engage different cell signaling pathways. Analyses of patient tissue reveal that PKAc-L205R mobilizes the Hippo signaling pathway to control organ size. Conversely phenotypic profiling of adrenal-specific knock-in mice indicate that PKAc-W196R activates the ERK mitogenic kinase cascade. Aim 2 will evaluate i) if female PKAcWT/W196R mice are more prone to Cushing’s comorbidities, ii) test if PKAc-L205R engages the Hippo signaling cascade to drive cortisol hyperproduction and iii) ascertain if PKAc-W196R coordinates ERK signaling events to promote adrenal hyperplasia. This proposal explores two transformative ideas that will change how we think about Cushing’s syndrome. 1) Losing spatiotemporal control of protein kinase A underlies adrenal Cushing’s pathologies. 2) Adjacent and identical mutations in PKAc drive adrenal hyperplasia and cortisol hypersecretion in Cushing’s patients.

抽象的 当肾上腺释放过多的胁迫同性皮质醇时，就会发生库欣综合症。这 内分泌障碍自然是由肾上腺或型肉体肿瘤引起的，妇女折磨了五倍 经常比男人。长期暴露于皮质醇还会增加血糖并升高血压。 该提案研究了肾上腺库欣综合征的分子发病机理。这种疾病通常是 肾上腺皮质中cAMP信号有缺陷的结果。精确医学的进步已经确定了点 蛋白激酶A催化亚基（PKAC）的突变和插入作为该疾病的驱动因素。最多 普遍的突变体是PKAC-L205R，在约45％的库欣患者中发现。其他遗传病变包括 PKAC-W196R。两种致病突变都存在于激酶结构域的区域内，该区域与 它的监管亚基。该蛋白质蛋白界面对于激酶活性自身抑制是必要的 通过与激酶锚定蛋白相关的PKA全酶的亚细胞靶向 （Akaps）。我们的生化，基于细胞和临床分析推断出携带PKAC-L205R的突变等位基因 或W196R足以使库欣患者的正常亚细胞作用部位置换激酶。这 导致了一个有效的假设，即失去对蛋白激酶A的空间时间控制是肾上腺库欣的基础 综合征。 AIM 1将测试i）是否恢复PKA锚定修改皮质醇释放，ii）确定哪个AKAP 协调皮质醇生物合成和iii）认为PKAC变体的“底物”的相对前提 或改变酶稳定性驱动库欣综合症。乍一看，PKAC-L205R和PKAC-W196R 突变体看起来几乎相同。然而，我们发现这些是身体上不同的激酶 参与不同的电池信号通路。对患者组织的分析表明，PKAC-L205R动员了河马 控制组织尺寸的信号通路。相反的肾上腺特异性敲孔小鼠的表型分析表明 PKAC-W196R激活ERK有丝分裂激酶级联反应。 AIM 2将评估i）如果女性pkacwt/w196r 小鼠更容易容易出现库欣的合并症，ii）测试PKAC-L205R是否将河马信号级联接合到 驱动皮质醇超生产和iii）确定PKAC-W196R是否坐标ERK信号事件以促进 肾上腺增生。该建议探讨了两个变革性的想法，这些想法将改变我们的看法 库欣综合症。 1）失去蛋白激酶A的时空控制肾上腺库欣的基础 病理。 2）PKAC驱动肾上腺增生和皮质醇过度分泌的相邻和相同的突变 在库欣的病人中。

项目成果

AKAP Signaling Islands: Venues for Precision Pharmacology.

• DOI: 10.1016/j.tips.2020.09.007
• 发表时间: 2020-12
• 期刊: Trends in pharmacological sciences
• 影响因子: 13.8
• 作者: Omar MH;Scott JD
• 通讯作者: Scott JD

Classification of Cushing's syndrome PKAc mutants based upon their ability to bind PKI

• DOI: 10.1042/bcj20230183
• 发表时间: 2023-06-01
• 期刊: BIOCHEMICAL JOURNAL
• 影响因子: 4.1
• 作者: Omar，Mitchell H.;Kihiu，Maryanne;Scott，John D.
• 通讯作者: Scott，John D.

Proximity biotinylation to define the local environment of the protein kinase A catalytic subunit in adrenal cells.

• DOI: 10.1016/j.xpro.2022.101992
• 发表时间: 2023-03-17
• 期刊: STAR protocols
• 作者: Omar MH;Lauer SM;Lau HT;Golkowski M;Ong SE;Scott JD
• 通讯作者: Scott JD

Mislocalization of protein kinase A drives pathology in Cushing's syndrome.

• DOI: 10.1016/j.celrep.2022.111073
• 发表时间: 2022-07-12
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Omar, Mitchell H.;Byrne, Dominic P.;Jones, Kiana N.;Lakey, Tyler M.;Collins, Kerrie B.;Lee, Kyung-Soon;Daly, Leonard A.;Forbush, Katherine A.;Lau, Ho-Tak;Golkowski, Martin;McKnight, G. Stanley;Breault, David T.;Lefrancois-Martinez, Anne-Marie;Martinez, Antoine;Eyers, Claire E.;Baird, Geoffrey S.;Ong, Shao-En;Smith, F. Donelson;Eyers, Patrick A.;Scott, John D.
• 通讯作者: Scott, John D.

CG-NAP/Kinase Interactions Fine-Tune T Cell Functions.

CG-NAP/激酶相互作用微调 T 细胞功能。

• DOI: 10.3389/fimmu.2019.02642
• 发表时间: 2019
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Verma,NavinKumar;Chalasani,MadhaviLathaSomaraju;Scott,JohnD;Kelleher,Dermot
• 通讯作者: Kelleher,Dermot