【正文】 節(jié)機體免疫功能和針對病毒本身兩個方面。 The antivirus gene therapy is very arrestive. Regulation of immunological function and direct treatment of virus are two ways of antivirus gene therapy. 3/20/2023 137 ? 前者主要是將細胞因子（干擾素、白介素）的基因導入免疫細胞，誘導機體對病毒的細胞和體液免疫應答。 Transduction of cytokine genes eg. interferon and interleukin to induces humoral and cellular immune responses against virus. 3/20/2023 138 后者 The latter： ? 1）誘導對病毒 RNA的降解 induce degradation of viral RNA ? 2）抑制病毒與宿主細胞結合 inhibit virus to contact with host cells ? 3）干擾病毒基因組的轉錄起始 interfere the transcription initiation of viral genome. 3/20/2023 139 ? 抑制病毒復制和蛋白合成 inhibit viral replication and protein synthesis ? RNA干擾 (RNA interference, RNAi) 3/20/2023 140 .干擾 RNA， RNAi ?哺乳動物中的研究 1999年才開始 ?被認為是最有前途的核酸藥物 ?《 Science》雜志將這一新發(fā)現(xiàn)評為2023 年度世界十大科學成就之首 . 3/20/2023 141 In 1990, RNA silencing was first observed in plants. This phenomena was called “ cosuppression” or “ PTGS” (posttranscriptional gene silencing). Plant Cell, van der Krol et al . 1990, 2(4): 291299 The history of siRNA 3/20/2023 142 ?a 起始階段 ：由于 RNA 病毒感染、轉座子轉錄和外源導入基因表達等原因 , 生物體細胞內(nèi)出現(xiàn)雙鏈 RNA (dsRNA) , dsRNA 在一種具有 RNase Ⅲ 樣活性的 dsRNA 特異性核酸內(nèi)切酶 (dsRNA specificendonuclease , Dicer) 作用下 , 被裂解成由正義和反義鏈組成的 21～23 nt 的干擾性小 RNA ( small interfering RNA , 或 short interferingRNAs , siRNA) . 3/20/2023 143 b 效應階段 ： 目前認為該階段存在兩種假說模式 , 一種假說模式是認為 siRNA 作為向導序列 , 與 RNA 誘導的沉默復合物 ( RNAinduced silencing plex , RISC) 的無活性前體結合 , 在 ATP 參與下 siRNA 雙鏈結構解旋誘導形成有活性的 RISC , 活性 RISC 隨即與同源性靶向 mRNA 結合 , 在與 siRNA 反義鏈配對區(qū)域的中部切斷同源性靶向 mRNA , 使其降解 , 從而干擾基因表達 , 這稱為內(nèi)源性核苷酸裂解(endonucleolytic cleavage) 3/20/2023 144 ?另一種假說模式為隨機降解性多聚酶鏈反應 ( random degradative PCR) ,即 siRNA 作為針對靶向 mRNA 特殊引物 , 在 RNA依賴性 RNA 多聚酶 ( RNA dependent RNA polymerase , RdRP) 的作用下以靶向mRNA 為模板合成靶向 mRNA 的互補鏈 , 使單鏈 mRNA 成為 dsRNA , 然后在 Dicer 酶的作用下被降解形成新的 siRNA , 新的siRNA 又進入上述過程 , 反復循環(huán)，從而形成信號擴增系統(tǒng) . 3/20/2023 145 Mechanism of Short interfering RNAs — posttranscriptional gene silencing. 3/20/2023 146 四、其他疾病的基因治療 Gene therapy of other disorders ? 人神經(jīng)退化疾病 human neurodegenerative diseases （如帕金森氏病 Parkinson39。s disease ) 3/20/2023 147 More than million Americans suffer from Parkinson39。s disease. There is no cure. But now there is a new treatment that may improve patients39。 quality of life. 3/20/2023 148 基因治療的前景與問題 Problems Prospects in gene therapy 20世紀 80年代發(fā)展起來的最具革命性的醫(yī)療技術。 One of the most revolutionary medical technologies has been developed since the 1980s. 3/20/2023 149 3/20/2023 150 存在一系列理論和技術問題，需要大量基礎研究和臨床實驗來解決。 There are a series of theoretical and technical problems to solve by making a lot of basic researches and clinic experiments . 3/20/2023 151 3/20/2023 152 ? 安全性 SAFETY 世界各國對基因治療產(chǎn)品、方法都采取嚴格措施監(jiān)控。 Every nation in the world adopts strict measures to supervise the products and methods of human gene therapy. 3/20/2023 153 兩年前在 University of Pennsylvania(賓夕法尼亞州 ), 一名 18歲男孩在基因治療時死亡。 美國的基因治療陷入全面低潮。 A sudden breakthrough SUSAN DENTZER: The death of the teenager, 18yearold Jesse Gelsinger, who suffered from a rare liver disorder, occurred during an experiment at the university of Pennsylvania. Gelsinger was injected with special viruses designed to carry healthy copies of a gene into his body. His father, Paul Gelsinger, told Senators today that his son39。s disease was actually under control, but that he agreed to participate in the clinical trial to help other sufferers from the disease. PAUL GELSINGER: Jesse was doing exceptionally well on his medications, and nothing should have prevented him from living a full and happy life. He believed, after discussions with representatives from Penn, that the worst that could happen in the trial would be that he would have flulike symptoms for a week. He was excited to help. SUSAN DENTZER: 17 other patients had preceded Jesse into the trial and were apparently treated with few ill effects. But Gelsinger39。s case quickly turned to tragedy soon after he was injected with genes carried by the viral messenger, known as a vector. PAUL GELSINGER: Less than 24 hours after they injected Jesse with the vector in the amount that only one other person had ever been given, Jesse39。s entire body had been reacting adversely. He went into a a before I could get to Philadelphia and see him, and died two days after my arrival, directly as a result of that gene therapy experiment. 2023年 12月，深圳，中國宣布基因治療的合法性 3/20/2023 154 ? 倫理性 Ethical aspects 確?；蛑委熂夹g真正用于解除患者病痛，造福于人類。 To ensure gene therapy technologies to be used in cure of disorders of human . 3/20/2023 155 技術性問題 :Technical problems ? 1)基因調(diào)控元件的選擇 Select for gene regulatory element ? 2)安全高效載體的構建 To construct safe and efficient vector systems 3/20/2023 156 ? 3)定向轉移技術 Techniques of directed transfers ? 4)合適的靶細胞 suitable target cells 3/20/2023 157 小 結 conclusions 3/20/2023 158 Thank you for your attention! 3/20/2023 159 靜夜四無鄰，荒居舊業(yè)貧。 , February 27, 2023 雨中黃葉樹，燈下白頭人。 23:26:3823:26:3823:262/27/2023 11:26:38 PM 1以我獨沈久，愧君相見頻。 :26:3823:26Feb2327Feb23 1故人江海別，幾度隔山川。 23:26:3823:26:3823:26Monday, February 27, 2023 1乍見翻疑夢，相悲各問年。 :26:3823:26:38February 27, 2023 1他鄉(xiāng)生白發(fā)，舊國見青山。 2023年 2月 27日星期一 下午 11時 26分 38秒 23:26: 1比不了得就不比，得不到的就不要。 2023年 2月 下午 11時 26分 :26February 27, 2023 1行動出成果，工作出財富。 2023年 2月 27日星期一 11時 26分 38秒 23:26:3827 February 2023 1做前，能夠環(huán)視四周；做時，你只能或者最好沿著以腳為起點的射線向前。 下午 11時 26分 38秒 下午 11時 26分 23:26: 沒有失敗，只有暫時停止成功！。 , February 27, 2023 很多事情努力了未必有結果，但是不努力卻什么改變也沒有。 23:26:3823:26:3823:262/27/2023 11:26:38 PM 1成功就是日復一日那一點點小小努力的積累。 :26:3823:26Feb2327Feb23 1世間成事，不求其絕對圓滿