探索microRNAs作為末期腎病相關合併症之診斷生物標誌物研究。

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吳坤霖(Kun-Lin Wu) 查詢紙本館藏

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系統生物與生物資訊研究所

論文名稱

探索microRNAs作為末期腎病相關合併症之診斷生物標誌物研究。
(The investigation of the microRNAs as the diagnostic biomarkers for the comorbidities in end-stage renal disease patients.)

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至系統瀏覽論文 (2029-6-30以後開放)

摘要(中)

慢性腎臟病是一世界性的健康問題，隨著腎功能逐漸下降至末期腎病，電解質不平衡、酸血症、體液過多、貧血及尿毒症等問題隨之發生，需實行腎臟替代療法以維持生命與生活品質。在腎臟替代療法中，腹膜透析為一方便經濟的治療模式，但在長期腹膜透析液或感染等刺激腹膜下，可能會發展致命性的包覆性腹膜硬化症。因此病早期症狀無特異性，故常會延遲診斷導致高的死亡率。另腎臟病也與心血管相關疾患息息相關，而冠狀動脈疾患相關症狀於血液透析族群中較為隱晦不明，非侵入性檢查其準確率亦有偏低的現象，致未能早期即時診斷治療。
腎臟替代療法雖然可維持末期腎病患者生命，但是相關併發症的困難診斷仍讓病人置身於不可挽救的狀況中。微型RNA參與在重要的基因轉錄調控中，其常被包裹於胞外小體、泌外體或其他結構中，可以穩定存在於體液中扮演著訊息傳遞的角色，且有文獻報導可以應用微型RNA於疾病偵測以期能早期準確診斷。
在此研究中，透過系統性的方法發現腹膜透析液及血液中的微型RNA分別在包覆性腹膜硬化症及冠心症發生時會產生有意義的改變。以包覆性腹膜硬化症來說，篩選出有變化的微型RNA，結合臨床表現，透過統計及機器學習模式發展預測模型，可有效的將診斷準確率提高。再進一步利用資料庫分析，發現這些特定的微型RNA與分子訊息調控相關，亦與特定藥物有關聯，上述發現指向微型RNA本身及相關藥物可能具有治療的濳力。另一方面，冠狀動脈疾患在血液透析病人常因不易診斷導致猝死，我們利用微型RNA在血液中表現量的變化，結合異常生化值，發展出較目前非侵入性診斷更為準確的冠狀動脈疾患預測模型，同時透過資料庫發現其與冠狀動脈疾病發生的因子及機轉相關。未來將透過進一步實驗了解微型RNA在包覆性腹膜硬化症及冠狀動脈疾患扮演了何種重要角色，參與疾病形成機轉及在治療方面的潜力。

摘要(英)

Chronic kidney disease (CKD) is a global health concern. As renal function gradually declines to end-stage renal disease (ESRD), issues such as electrolyte imbalance, acidosis, fluid overload, anemia, and uremia develop, necessitating renal replacement therapy to sustain life and maintain life quality. Peritoneal dialysis (PD) is an economically convenient renal replacement therapy. However, prolonged exposure to PD fluid or infections may lead to the encapsulating peritoneal sclerosis (EPS). Early symptoms of this disease are uncertain, often resulting in delayed diagnosis with high mortality rates.
Additionally, the renal failure is closely associated with cardiovascular disease. Symptoms related to coronary artery disease in the hemodialysis (HD) population may be subtle, and the accuracy of non-invasive diagnostic tools is often limited, leading to delayed diagnoses and treatment.
While renal replacement therapy can sustain the lives in end-stage renal disease patients, the challenging diagnosis of complications places patients at an irretrievable high risk of mortality. MicroRNA (miRNA) plays a crucial role in regulating gene transcription. Encased in extracellular vesicles, exosomes, or other structures in circulation, miRNAs stably exist in biofluids, serving as messengers. Literatures report that the miRNAs can be applied as biomarkers for disease detection.
In this study, a systematic approach revealed significant changes in miRNA expressions in PD effluent and blood of EPS and CAD, respectively. For EPS, altered miRNAs were identified, and, when combined with clinical manifestations, a predictive model was developed using statistical and machine learning models to improve diagnostic accuracy. Further database analysis indicated that these candidate miRNAs are associated with molecular message pathway and linked to specific drugs, suggesting therapeutic potential for both microRNA itself and related drugs.
On the other hand, CAD in HD patients often leads to sudden death due to the difficulty in diagnosis. By utilizing variations in miRNA expression in blood, coupled with abnormal biochemical values, a more accurate predictive model for CAD was developed compared to current non-invasive diagnostics. Database findings revealed these miRNAs involved pathophysiology. Future experiments will explore the critical roles that miRNAs play in EPS and CAD, elucidating the mechanisms of disease and the therapeutic potential.

關鍵字(中)

★ 微型RNA
★ 末期腎病
★ 腹膜透析
★ 血液透析
★ 包覆性腹膜硬化症
★ 冠狀動脈疾患

關鍵字(英)

★ microRNA
★ end-stage renal disease
★ peritoneal dialysis
★ hemodialysis
★ encapsulating peritoneal sclerosis

論文目次

目錄
中文摘要 I
ABSTRACT III
ACKNOWLEDGEMENT V
LIST OF FIGURES XI
LIST OF TABLES XIII
LIST OF ABBREVIATIONS XIV
CHAPTER 1: LITERATURE REVIEW 1
1.1. INTRODUCTION TO CHRONIC KIDNEY DISEASE 1
1.1.1 Renal function 1
1.1.2 Chronic kidney disease 7
1.1.3 Renal replacement therapy 9
1.2 INTRODUCTION TO ENCAPSULATING PERITONEAL SCLEROSIS. 11
1.2.1 Signs and symptoms of encapsulating peritoneal sclerosis 12
1.2.2 Diagnosis for encapsulating peritoneal sclerosis. 13
1.2.3 The treatment of encapsulating peritoneal sclerosis 15
1.3 INTRODUCTION TO CORONARY ARTERY DISEASE IN HEMODIALYSIS PATIENTS 16
1.3.1 Signs and symptoms of coronary artery disease 16
1.3.2 Coronary artery disease in hemodialysis patients 18
1.3.3 The diagnosis for coronary artery disease 19
1.4 INTRODUCTION TO MICRORNA 20
1.4.1 Biomarkers for diseases 20
1.4.2 Biogenesis and mechanism of microRNA 21
1.4.3 microRNA dysregulation in disease 22
1.5 SIGNIFICANCES AND PURPOSE 23
CHAPTER 2. MATERIALS AND METHODS 32
2.1 PATIENT RECRUITMENTS WITH ETHICS APPROVAL AND INFORMED CONSENT 32
2.2 PREPARATION OF PD EFFLUENT AND PLASMA AND MIRNA ISOLATION AND QUANTIFICATION BY RT-PCR 33
2.3 HIGH-THROUGHPUT AND SINGLE REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION ANALYSIS 35
2.4 NEXT GENERATION SEQUENCE OF MIRNAS 36
2.5 MULTIPLE LOGISTIC REGRESSION AND MACHINE LEARNING MODEL 37
2.6 THE CONSTRUCTION OF MIRNA-GENE INTERACTION NETWORK AND PATHWAY ENRICHMENT ANALYSIS 38
2.7 THE CONSTRUCTION OF MIRNA-COMPOUND INTERACTION NETWORK 38
2.8 STATISTICAL ANALYSIS 39
CHAPTER 3. THE MIRNA CANDIDATE AS DIAGNOSTIC TOOL FOR ENCAPSULATING PERITONEAL SCLEROSIS IN PERITONEAL DIALYSIS PATIENTS. 40
3.1 INTRODUCTION 40
3.2 RESULTS 42
3.2.3 Identification of candidate miRNAs in effluent between EPS and non-EPS 42
3.2.4 The predication model to identify EPS in PD patients. 44
3.2.5 The targeting genes of EPS associated miRNAs and the pathways. 46
3.2.6 The interactions of EPS associated miRNAs and drug compounds. 47
3.3 DISCUSSION 48
CHAPTER 4. CANDIDATE MIRNAS AS DETECTORS FOR CORONARY ARTERY DISEASE IN HEMODIALYSIS PATIENTS 70
4.1 INTRODUCTION 70
4.2 RESULTS 71
4.2.1 The clinical characteristics in hemodialysis patients with coronary artery. 71
4.2.2 Identification of candidate miRNAs of plasma in hemodialysis patients with coronary artery. 72
4.2.3 The miRNA based predication model of coronary artery disease in hemodialysis patients. 73
4.2.5 The interactions of CAD associated miRNAs and drug compounds. 75
4.3 DISCUSSION 76
CHAPTER 5. CONCLUDING REMARKS AND FUTURE DIRECTION 94
5.1 CONCLUSION REMARKS 94
5.2 FUTURE DIRECTION 97
5.2.1 To boost ability of prediction model for more accuracy and early detection. 97
5.2.2 To investigate the interplay of miRNAs and pathophysiology of EPS. 97
5.2.3 To investigate the interaction between miRNAs and vascular disease in HD population. 99
101
REFERENCES 105
APPENDIX A. PUBLICATION LIST. 117

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指導教授

馬念涵(Nianhan Ma)

審核日期

2024-5-22

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