Innovation Center for Neurological Disorders and Department of Neurology, Xuanwu Hospital, Capital Medical University, National Clinical Research Center for Geriatric Diseases, Beijing 100053, China.
Department of Psychiatry, Washington University, St. Louis, MO 63110, USA.
Hum Mol Genet. 2021 May 28;30(9):811-822. doi: 10.1093/hmg/ddab090.
To identify novel risk genes and better understand the molecular pathway underlying Alzheimer's disease (AD), whole-exome sequencing was performed in 215 early-onset AD (EOAD) patients and 255 unrelated healthy controls of Han Chinese ethnicity. Subsequent validation, computational annotation and in vitro functional studies were performed to evaluate the role of candidate variants in EOAD. We identified two rare missense variants in the phosphodiesterase 11A (PDE11A) gene in individuals with EOAD. Both variants are located in evolutionarily highly conserved amino acids, are predicted to alter the protein conformation and are classified as pathogenic. Furthermore, we found significantly decreased protein levels of PDE11A in brain samples of AD patients. Expression of PDE11A variants and knockdown experiments with specific short hairpin RNA (shRNA) for PDE11A both resulted in an increase of AD-associated Tau hyperphosphorylation at multiple epitopes in vitro. PDE11A variants or PDE11A shRNA also caused increased cyclic adenosine monophosphate (cAMP) levels, protein kinase A (PKA) activation and cAMP response element-binding protein phosphorylation. In addition, pretreatment with a PKA inhibitor (H89) suppressed PDE11A variant-induced Tau phosphorylation formation. This study offers insight into the involvement of Tau phosphorylation via the cAMP/PKA pathway in EOAD pathogenesis and provides a potential new target for intervention.
为了鉴定新的风险基因,并更好地了解阿尔茨海默病(AD)的分子途径,我们对 215 名早发性 AD(EOAD)患者和 255 名汉族无关健康对照者进行了全外显子测序。随后进行了验证、计算注释和体外功能研究,以评估候选变异在 EOAD 中的作用。我们在 EOAD 个体的磷酸二酯酶 11A(PDE11A)基因中发现了两个罕见的错义变异。这两个变异都位于进化上高度保守的氨基酸中,预计会改变蛋白质构象,并被归类为致病性的。此外,我们发现 AD 患者的大脑样本中 PDE11A 的蛋白水平显著降低。PDE11A 变异的表达和针对 PDE11A 的特异性短发夹 RNA(shRNA)的敲低实验都导致 Tau 在多个表位的 AD 相关过度磷酸化增加。PDE11A 变异或 PDE11A shRNA 也导致 cAMP 水平、蛋白激酶 A(PKA)激活和 cAMP 反应元件结合蛋白磷酸化增加。此外,PKA 抑制剂(H89)预处理抑制了 PDE11A 变异诱导的 Tau 磷酸化形成。这项研究深入了解了 Tau 磷酸化通过 cAMP/PKA 途径在 EOAD 发病机制中的作用,并为干预提供了一个潜在的新靶点。