基于生物信息學(xué)對耐藥結(jié)核分枝桿菌潛在藥物靶點的挖掘分析
發(fā)布時間：2018-08-31 13:09
【摘要】：結(jié)核病是全球性的重大健康問題,近年來,由于艾滋病病毒與結(jié)核分枝桿菌雙重感染的流行,加之抗生素的濫用,使結(jié)核分枝桿菌多重耐藥問題日趨嚴(yán)峻,已成為結(jié)核病防治的重大難題,也是結(jié)核病全球蔓延、疫情惡化的重要原因。自1963年發(fā)現(xiàn)利福平、異煙肼等一線抗結(jié)核藥物以來,尚未研發(fā)出新型的、高效的抗結(jié)核藥物用以替代傳統(tǒng)的抗結(jié)核藥物。究其原因,主要是由于結(jié)核分枝桿菌有效的藥物作用靶分子數(shù)量不足,遠遠不能滿足當(dāng)前結(jié)核病防治的需要,尤其是多重耐藥結(jié)核分枝桿菌治療的需要。因此結(jié)核分枝桿菌藥物作用靶標(biāo)的高通量篩選是新型抗結(jié)核藥物研發(fā)及結(jié)核病防治的第一步,也是關(guān)鍵的一步。結(jié)核分枝桿菌細胞壁主要由莢膜、分支菌酸、肽聚糖、阿拉伯糖、內(nèi)膜組成。這些結(jié)構(gòu)對于維持結(jié)核分枝桿菌細胞形態(tài)的完整性、抵御外界化學(xué)物質(zhì)的侵蝕、逃脫免疫以及產(chǎn)生耐藥性和致病性都起著重要的作用,因此結(jié)核分枝桿菌細胞壁相關(guān)物質(zhì)代謝及合成途徑可以作為抗結(jié)核藥物作用靶點。傳統(tǒng)的抗結(jié)核菌一線藥物如異煙肼等,其作用靶點也正是抑制結(jié)核分枝桿菌細胞壁中的分枝菌酸的合成。但是結(jié)核分枝桿菌的基因型極其復(fù)雜和多變,傳統(tǒng)的藥物作用靶點如inhA、ahpC、nadh、katG、KasA等都出現(xiàn)了耐藥性突變,新的細胞壁相關(guān)基因的篩選迫在眉睫。此外,結(jié)核分枝桿菌細胞壁的組成成分復(fù)雜、功能繁多,傳統(tǒng)的結(jié)核分枝桿菌細胞壁相關(guān)基因的篩選主要通過基因或敲除或RNA沉默等方法對潛在的細胞壁合成及代謝靶分子進行逐一的篩選,實驗成本高,效率低,而且由于實驗技術(shù)的限制,目前尚無任何一種實驗方法可以實現(xiàn)全基因組的細胞壁合成相關(guān)分子的系統(tǒng)性篩選,因此,亟待建立以結(jié)核分枝桿菌信號傳導(dǎo)動態(tài)網(wǎng)絡(luò)為基礎(chǔ)的細胞壁合成相關(guān)基因全景掃描及篩選新模式。鑒于此,本研究收集了2013年5月之前的所有結(jié)核分枝桿菌H37Rv的基因芯片共計43個系列2861張芯片。通過整合聚類,建立結(jié)核分枝桿菌細胞壁相關(guān)基因、未知基因和一般基因的三元素網(wǎng)絡(luò)圖,通過聚類基因簇的模體分析,高通量注釋結(jié)核分枝桿菌細胞壁相關(guān)基因,為研發(fā)有效的、敏感的、副作用低的抗結(jié)核藥物奠定分子基礎(chǔ)。到目前為止人們關(guān)于異煙肼對結(jié)核分枝桿菌的影響研究結(jié)果仍然表淺,公認的結(jié)論僅僅停留在其對結(jié)核分枝桿菌的KatG基因和對細胞壁的結(jié)構(gòu)產(chǎn)生影響這一層面。根據(jù)人們對該基因的功能研究和細菌形態(tài)學(xué)發(fā)現(xiàn)其主要功能都集中于細菌耐藥的相關(guān)研究,但對于異煙肼如何殺傷細菌的機制研究甚少。異煙肼一定對分枝桿菌產(chǎn)生了更致命的作用,但這些作用由于觀察和證實的困難性還沒有被揭示。而這往往才是最主要的作用。對于如何揭示這些藥物作用靶點,我們考慮利用藥物的副作用作為切入點來探索藥物對分枝桿菌的作用靶點。藥物作用于人體后產(chǎn)生一系列的生物化學(xué)反應(yīng),人們利用藥物主要反應(yīng)治療疾病,并且盡量規(guī)避眾多副作用。但是就是這些副作用為確定藥物作用靶點提供了可能。Monica Campillos等人通過不同藥物相似的副反應(yīng)推斷他們有相同的作用靶點。這個理論為我們探索藥物作用靶點提供了可能。異煙肼是一線抗結(jié)核藥物,在治療結(jié)核病的同時也會對人體的肝臟、維生素B6的代謝和氧化應(yīng)激過程產(chǎn)生影響。這也證明這些組織和生物過程中共同存在著異煙肼的結(jié)合靶點或作用靶點。而且異煙肼又是可以作用于結(jié)核的藥物,那么人體和結(jié)核分枝桿菌中一定存在著一個或一些異煙肼共同作用的靶點。聯(lián)合分析異煙肼作用于結(jié)核分枝桿菌和人體的相關(guān)基因,通過大量交互性基因序列比對找到共同的基因序列,從生物學(xué)和統(tǒng)計學(xué)的角度進一步分析其成為藥物作用靶點的可能性。這種方法突破了傳統(tǒng)的細胞學(xué)或者分子學(xué)方法篩選藥物作用靶點的瓶頸,極大的提高篩選靶點的速度和效率。為研究異煙肼的藥理及對結(jié)核分枝桿菌細菌的致死作用提供了理論基礎(chǔ)。
[Abstract]:Tuberculosis is a major global health problem. In recent years, due to the epidemic of double infection of HIV and Mycobacterium tuberculosis, coupled with the abuse of antibiotics, the problem of multiple drug resistance of Mycobacterium tuberculosis has become increasingly serious. It has become a major problem in tuberculosis control and prevention. It is also an important reason for the global spread of tuberculosis and the deterioration of the epidemic situation. Since the discovery of rifampicin, isoniazid and other first-line anti-tuberculosis drugs in 1997, no new, highly effective anti-tuberculosis drugs have been developed to replace traditional anti-tuberculosis drugs. The main reason is that the number of effective drug targets of Mycobacterium tuberculosis is insufficient, far from meeting the needs of current tuberculosis control, especially multiple anti-tuberculosis drugs. High-throughput screening of targets for drug-resistant Mycobacterium tuberculosis is therefore the first and key step in the development of new anti-tuberculosis drugs and in the prevention and treatment of tuberculosis. Mycobacterium tuberculosis cell morphological integrity, resistance to external chemical erosion, escape from immunity and the production of drug resistance and pathogenicity play an important role, so the metabolism and synthesis of cell wall-related substances of Mycobacterium tuberculosis can be used as a target of anti-tuberculosis drugs. Traditional first-line anti-tuberculosis drugs such as isoniazid However, the genotype of Mycobacterium tuberculosis is extremely complex and changeable. The traditional drug targets such as inhA, ahpC, nadh, katG, KasA, etc. all have drug resistance mutations, and the screening of new cell wall-related genes is imminent. The components of cell wall of Mycobacterium tuberculosis are complex and have many functions. The traditional screening methods of genes related to cell wall of Mycobacterium tuberculosis mainly through gene knockout or RNA silencing to screen the potential target molecules of cell wall synthesis and metabolism one by one. The experimental cost is high and the efficiency is low. Therefore, it is urgent to establish a new panoramic scanning and screening model for genes related to cell wall synthesis based on the dynamic signal transduction network of Mycobacterium tuberculosis. Bacillus H37Rv gene chips consist of 43 series of 2861 chips. Through integrated clustering, a three-element network diagram of cell wall-related genes, unknown genes and general genes of Mycobacterium tuberculosis was established. Through the analysis of clustering gene clusters, high-throughput annotation of cell wall-related genes of Mycobacterium tuberculosis was made, which is an effective, sensitive and side-effect for research and development. The molecular basis is laid by low levels of anti-tuberculosis drugs. Up to now, studies on the effect of isoniazid on Mycobacterium tuberculosis are still superficial, and the accepted conclusion is only that it affects the KatG gene of Mycobacterium tuberculosis and the structure of the cell wall. Morphological studies have found that the main functions of isoniazid are related to bacterial resistance, but little is known about how isoniazid kills bacteria. Isoniazid must have a more lethal effect on Mycobacterium, but these effects have not been revealed due to the difficulty of observation and confirmation. This is often the most important role. To uncover these drug targets, we consider using the side effects of drugs as a starting point to explore the targets of drugs on Mycobacterium. Drugs act on the human body and produce a series of biochemical reactions. People use the main reactions of drugs to treat diseases and try to avoid many side effects. But these side effects are Monica Campillos et al. inferred that different drugs had the same target through similar side effects. This theory provides a possibility for us to explore drug targets. Isoniazid is a first-line anti-tuberculosis drug that treats tuberculosis but also generates vitamin B6 in the human liver. It also proves that these tissues and biological processes have the binding or action targets of isoniazid, and isoniazid is a drug that can act on tuberculosis, so there must be one or some targets of isoniazid in human and Mycobacterium tuberculosis. Nicotinic hydrazine acts on the related genes of Mycobacterium tuberculosis and human body, finds the common gene sequence through a large number of interactive gene sequence alignment, and further analyzes the possibility of nicotinic hydrazine as a drug target from the biological and statistical point of view. This study provides a theoretical basis for studying the pharmacology of isoniazid and its lethal effect on Mycobacterium tuberculosis.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R96
本文編號：2215082
[Abstract]:Tuberculosis is a major global health problem. In recent years, due to the epidemic of double infection of HIV and Mycobacterium tuberculosis, coupled with the abuse of antibiotics, the problem of multiple drug resistance of Mycobacterium tuberculosis has become increasingly serious. It has become a major problem in tuberculosis control and prevention. It is also an important reason for the global spread of tuberculosis and the deterioration of the epidemic situation. Since the discovery of rifampicin, isoniazid and other first-line anti-tuberculosis drugs in 1997, no new, highly effective anti-tuberculosis drugs have been developed to replace traditional anti-tuberculosis drugs. The main reason is that the number of effective drug targets of Mycobacterium tuberculosis is insufficient, far from meeting the needs of current tuberculosis control, especially multiple anti-tuberculosis drugs. High-throughput screening of targets for drug-resistant Mycobacterium tuberculosis is therefore the first and key step in the development of new anti-tuberculosis drugs and in the prevention and treatment of tuberculosis. Mycobacterium tuberculosis cell morphological integrity, resistance to external chemical erosion, escape from immunity and the production of drug resistance and pathogenicity play an important role, so the metabolism and synthesis of cell wall-related substances of Mycobacterium tuberculosis can be used as a target of anti-tuberculosis drugs. Traditional first-line anti-tuberculosis drugs such as isoniazid However, the genotype of Mycobacterium tuberculosis is extremely complex and changeable. The traditional drug targets such as inhA, ahpC, nadh, katG, KasA, etc. all have drug resistance mutations, and the screening of new cell wall-related genes is imminent. The components of cell wall of Mycobacterium tuberculosis are complex and have many functions. The traditional screening methods of genes related to cell wall of Mycobacterium tuberculosis mainly through gene knockout or RNA silencing to screen the potential target molecules of cell wall synthesis and metabolism one by one. The experimental cost is high and the efficiency is low. Therefore, it is urgent to establish a new panoramic scanning and screening model for genes related to cell wall synthesis based on the dynamic signal transduction network of Mycobacterium tuberculosis. Bacillus H37Rv gene chips consist of 43 series of 2861 chips. Through integrated clustering, a three-element network diagram of cell wall-related genes, unknown genes and general genes of Mycobacterium tuberculosis was established. Through the analysis of clustering gene clusters, high-throughput annotation of cell wall-related genes of Mycobacterium tuberculosis was made, which is an effective, sensitive and side-effect for research and development. The molecular basis is laid by low levels of anti-tuberculosis drugs. Up to now, studies on the effect of isoniazid on Mycobacterium tuberculosis are still superficial, and the accepted conclusion is only that it affects the KatG gene of Mycobacterium tuberculosis and the structure of the cell wall. Morphological studies have found that the main functions of isoniazid are related to bacterial resistance, but little is known about how isoniazid kills bacteria. Isoniazid must have a more lethal effect on Mycobacterium, but these effects have not been revealed due to the difficulty of observation and confirmation. This is often the most important role. To uncover these drug targets, we consider using the side effects of drugs as a starting point to explore the targets of drugs on Mycobacterium. Drugs act on the human body and produce a series of biochemical reactions. People use the main reactions of drugs to treat diseases and try to avoid many side effects. But these side effects are Monica Campillos et al. inferred that different drugs had the same target through similar side effects. This theory provides a possibility for us to explore drug targets. Isoniazid is a first-line anti-tuberculosis drug that treats tuberculosis but also generates vitamin B6 in the human liver. It also proves that these tissues and biological processes have the binding or action targets of isoniazid, and isoniazid is a drug that can act on tuberculosis, so there must be one or some targets of isoniazid in human and Mycobacterium tuberculosis. Nicotinic hydrazine acts on the related genes of Mycobacterium tuberculosis and human body, finds the common gene sequence through a large number of interactive gene sequence alignment, and further analyzes the possibility of nicotinic hydrazine as a drug target from the biological and statistical point of view. This study provides a theoretical basis for studying the pharmacology of isoniazid and its lethal effect on Mycobacterium tuberculosis.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R96
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