東亞特有乙醛去氫酶及葡萄糖六磷酸去氫酶基因變異與表型相關:串聯健保資料庫和台灣人體生物資料庫

Abstract

乙醛去氫酶ALDH2 (acetaldehyde dehydrogenase 2) 為粒線體中的一種酵素, 可幫助人體代謝飲用酒精後產生的的乙醛（acetaldehyde），以及許多體內產生與環境中的有毒醛類如4-hydroxynonenal (4-HNE)等，使其成為不具毒性之代謝物幫助排出人體。而這些對人體有害的醛類，會藉由發動親電子基攻擊蛋白質產生共價鍵結合，造成蛋白質失去功能。東亞有高達~ 40%的居民(約5.6 億)攜帶特有的ALDH2失活性點突變(rs671, Glu487Lys)，此基因突變為全球盛行率最高之單一基因疾病 (占~8% 全球人口)。 而另外一種亦為世界上最常見的酵素疾病則是G6PD(glucose-6-phosphate dehydrogenase)，全球約有3至4億人攜帶至少一個缺陷的G6PD(glucose-6-phosphate dehydrogenase, G6PD)基因變異，使得G6PD基因突變成為世界第二常見的單基因突變。在東亞以c.1376G>T(p.R459L)的點突變(廣東型蠶豆症)最常見。位置在Ｘ染色體遠端長臂 (Xq28）。在台灣，G6PD基因變異患病率約為3-5%，其中以c.1376G>T(p.R459L)的點突變(廣東型蠶豆症）最常見，占46.8%。。 缺乏glucose-6-phosphate dehydrogenase使細胞無法產出足夠的nicotinamide adeninedinucleotide phosphate (NADPH)，伴隨無法產生足夠reduced glutathione (GSH)，使氧化壓力增加。傳統觀念認為蠶豆症只會引起溶血或嚴重時產生黃疸, 但全身氧化壓力的增加，很可能造成其他病變，只是因為病患多局部分布於東南亞國家，所以研究不多。這些攜帶者的自然病程尚不清楚，其是否易受氧化壓力相關的退化性疾病（degenerative disease）影響也仍然未知。 我們實驗室先前的研究發現仿人類 Glu504Lys 突變的 Aldh2 knock‑in mouse由於適應性產熱和能量消耗減少，容易出現飲食引起的肥胖、葡萄糖不耐症、胰島素抗性和脂肪肝。我們亦發現模擬東亞廣東型蠶豆症突變位點之基因嵌入鼠會產生腎功能異常，包含遠端腎小管壞死以及心肌肥大，與肌少症。基於此發現，我們推測為氧化壓力增加，造成之腎傷害與心臟病變。基於G6PD促進劑的快速發展，以及治療慢性腎病病變，主要心臟不良事件(Major Adverse Cardiovascular Events)與肌少症的藥物治療有限，確認蠶豆症在人類會引發腎傷害與主要心臟不良事件有其必要。 而在臨床方面,我國由民國73年開始全國新生兒先天代謝異常疾病(包含蠶豆症),篩檢率達99%以上，累計篩檢超過7百萬名新生兒，其中以蠶豆症最多近12萬人，由於篩檢技術精準，使得台灣的蠶豆症診斷登錄精確而完整。 基於此初步結果，我們 1. 將台灣人體生物資料庫203,021的基因型，串連衛生福利部衛生福利資料科學中心台灣全民健保資料庫，探討G6PD基因突變與ALDH2基因變異對於退化性方面疾病:阿滋海默症、帕金森氏症、肌少症以及糖尿病共病方面疾病:第二型糖尿病、肥胖、慢性腎病病變、糖尿病神經病變、糖尿病視網膜病變，與主要心臟不良事件裡面心肌病變、心血管退化疾病例如:急性心肌梗塞、心臟衰竭、中風、心肌血管重建的相關。 2. 在衛生福利部衛生福利資料科學中心以多變量邏輯式迴歸以及傾向分數一對一的配對及加權，分析帶有rs671 rs72554664 rs72554665變異位點的人口中，罹患阿滋海默症、帕金森氏症、肌少症、肥胖、第二型糖尿病、慢性腎病病變、糖尿病神經病變、糖尿病視網膜病變、與重大心血管不良事件相關的比率。 在本研究中，於串聯台灣人體生物資料庫與衛生福利部衛生福利資料科學中心資料庫中整合基因型與臨床資料，並以多變量邏輯式迴歸以及傾向分數加權及配對分析，探討 rs671、rs72554664 (Kaiping)、rs72554665 (Canton) 與多種疾病的關聯。結果顯示：rs671 與阿茲海默症存在顯著相關；rs72554664 (Kaiping) 與第二型糖尿病存在顯著相關；rs72554665 (Canton) 則與慢性腎臟病變及腦中風存在顯著相關。在所有模型中皆依照不同疾病特性納入臨床共變數進行調整，以降低潛在混雜影響。整體而言，三個位點分別在四種疾病上觀察到罹患疾病與否存在顯著差異，顯示特定位點變異對多重疾病風險具有潛在貢獻。

Acetaldehyde dehydrogenase 2 (ALDH2) is a mitochondrial enzyme that detoxifies acetaldehyde, the metabolite generated from alcohol consumption, as well as other toxic endogenous and environmental aldehydes such as 4-hydroxynonenal (4-HNE). These harmful aldehydes can covalently bind to proteins via electrophilic attack, leading to loss of protein function. Approximately 40% of East Asians (~560 million individuals) carry the inactive ALDH2 point mutation rs671 (Glu487Lys), making it the most prevalent single-gene defect worldwide (~8% of the global population). Another common enzymopathy is glucose-6-phosphate dehydrogenase (G6PD) deficiency, with an estimated 300–400 million carriers globally, rendering G6PD mutations the second most frequent single-gene defects worldwide. In East Asia, the most common variant is the point mutation c.1376G>T (p.R459L), also known as the Canton type, located on the distal long arm of the X chromosome (Xq28). In Taiwan, the prevalence of G6PD mutations is about 3–5%, with the c.1376G>T variant being the most frequent, accounting for 46.8% of cases. Deficiency of G6PD leads to impaired production of nicotinamide adenine dinucleotide phosphate (NADPH), resulting in insufficient reduced glutathione (GSH) and increased oxidative stress. Traditionally, G6PD deficiency is thought to cause only hemolysis or neonatal jaundice in severe cases. However, systemic oxidative stress may also contribute to other pathologies. Due to the geographic distribution of patients being concentrated in Southeast Asia, research remains limited, and the natural history of these carriers is still unclear, particularly regarding their susceptibility to oxidative stress–related degenerative diseases. Our previous studies demonstrated that Aldh2 knock-in mice carrying the humanized Glu504Lys mutation exhibited reduced adaptive thermogenesis and energy expenditure, leading to susceptibility to diet-induced obesity, glucose intolerance, insulin resistance, and hepatic steatosis. In our animal model, mice with the East Asian Canton-type G6PD mutation exhibited renal dysfunction, including distal tubular necrosis, cardiac hypertrophy, and sarcopenia. These findings suggest that increased oxidative stress induces kidney injury and cardiac pathology. Given the rapid development of G6PD activators and the limited therapeutic options for chronic kidney disease (CKD), major adverse cardiovascular events (MACE), and sarcopenia, clarifying whether G6PD deficiency predisposes humans to kidney injury and cardiovascular events is essential. Clinically, Taiwan has implemented nationwide newborn screening for inherited metabolic disorders (including G6PD deficiency) since 1984, with a coverage rate exceeding 99% and more than 7 million newborns screened. Among them, G6PD deficiency is the most common, with nearly 120,000 identified cases. The precision of newborn screening ensures that the registry of G6PD deficiency in Taiwan is both accurate and comprehensive. To integrate 203,021 genotypes from the Taiwan Biobank with the National Health Insurance Research Database (NHIRD) at the Health and Welfare Data Science Center (HWDC), in order to examine associations between G6PD mutations and ALDH2 variants with degenerative diseases (Alzheimer’s disease, Parkinson’s disease, sarcopenia) and comorbidities of diabetes (type 2 diabetes, obesity, CKD, diabetic neuropathy, diabetic retinopathy), as well as major adverse cardiovascular events (including cardiomyopathy, acute myocardial infarction, heart failure, stroke, and coronary revascularization). To apply propensity score 1:1 matching and weighting analyses within the HWDC, evaluating the incidence of Alzheimer’s disease, Parkinson’s disease, sarcopenia, obesity, type 2 diabetes, CKD, diabetic neuropathy, diabetic retinopathy, and major adverse cardiovascular events among individuals carrying rs671, rs72554664, and rs72554665 variants. In observational studies, data are often subject to bias and confounding. Propensity score matching helps reduce such issues, allowing for more balanced comparisons between exposed and unexposed groups. In our analysis, the treatment variable was defined as the presence of specific genetic variants (rs671, rs72554664, rs72554665), and covariates were included in logistic regression models to estimate the propensity score for each subject. In PSM with 1:1 matching, each “treated” subject (variant carrier) is paired with one “control” subject (non-carrier), making the groups comparable in terms of age, sex, blood pressure, blood glucose, and other covariates, thereby approximating the conditions of a randomized controlled trial. In this study, by linking the Taiwan Biobank and NHIRD, we integrated genotypes with clinical data and applied propensity score weighting and matching to assess the associations of rs671, rs72554664 (Kaiping), and rs72554665 (Canton) with multiple diseases. Our results demonstrated that rs671 was significantly associated with Alzheimer’s disease, rs72554664 (Kaiping) with obesity, and rs72554665 (Canton) with CKD, stroke, and diabetic peripheral neuropathy. Disease-specific covariates such as HbA1c were included in the models to reduce potential confounding. Overall, significant associations were observed for the three loci across five diseases, suggesting their potential contributions to disease susceptibility. For future directions, we plan to conduct experiments using animal models to simulate the East Asian Canton-type G6PD mutation and evaluate its role in stroke susceptibility. Furthermore, our team intends to extend investigations to alcohol-metabolizing genes (particularly ALDH2 rs671 and ADH1B rs1229984). By combining genetic and lifestyle data from the Taiwan Biobank with clinical outcomes from the NHIRD, we aim to clarify the contributions of genotype and alcohol consumption to major chronic diseases (e.g., Alzheimer’s disease, Parkinson’s disease, type 2 diabetes, heart failure, CKD, obesity, sarcopenia). These results may also provide evidence for policy research, such as evaluating the potential health and social benefits of alcohol taxation in Taiwan.