【正文】 Somatic gene therapy 3/20/2023 122 選擇轉(zhuǎn)移基因的靶細(xì)胞，要考慮幾點(diǎn)： How to select target cells for gene transfer? ? 1）最好是組織 特異性 細(xì)胞 Tissue specific cells are better. ? 2）細(xì)胞較易獲得， It is easy to obtain. ? 3) 生命周期較長 Lifecycle is long. 3/20/2023 123 4）易受外源物質(zhì)轉(zhuǎn)化 It is easy to be transform by foreign factors 5）轉(zhuǎn)染后細(xì)胞仍易成活 It is easy to survive after transfection. 3/20/2023 124 目前已開展的研究的靶細(xì)胞有： Target cells in present studies 造血干細(xì)胞 (hematopoietic stem cells )、淋巴細(xì)胞 (lymphocytes)、皮膚成纖維細(xì)胞 (skin fibroblasts)、肝細(xì)胞 (hepatocytes)、血管內(nèi)皮細(xì)胞 (vascular endothelial cells,)、肌細(xì)胞 (muscular cells, )、神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞 (neurons glial cells )、腫瘤細(xì)胞 (tumour cells ) 等 and so on. 3/20/2023 125 ex vivo retroviral vectors (oncoretroviral or lentiviral) in vivo adenoviral vectors AAV vectors lentiviral vectors 3/20/2023 126 第三節(jié) 基因治療的應(yīng)用 applications of gene therapy ? 1990年 基因治療的第一個(gè)病例是先天性腺苷脫氨酶 (ADA)缺乏癥 The first disease approved for treatment with gene therapy was adenosine deaminase (ADA) deficiency in 1990. 3/20/2023 127 美國醫(yī)學(xué)家 W這個(gè) 4歲女孩由于遺傳基因有缺陷，自身不能生產(chǎn) ADA，先天性免疫功能不全，只能生活在無菌的隔離帳里。 謝德爾， 1999 3/20/2023 128 一、對(duì)遺傳病進(jìn)行基因治療 Gene therapy of human geic disease 研究范圍十分有限，須符合以下條件： The range of research is very limited, which must match the following conditions. 1）在 DNA水平上明確發(fā)病原因及機(jī)制 To define the causes and mechanisms of diseases at the DNA level. 3/20/2023 129 2）必須是單基因遺傳病 It must be a monogeic or single －gene disease. 3/20/2023 130 3）表達(dá)不需要精確調(diào)控 Accurate regulation of gene expression is not essential. 4）該遺傳病不經(jīng)治療有嚴(yán)重的后果 Severe results may be got without treatment. 3/20/2023 131 二、對(duì)惡性腫瘤的基因治療 Cancer Gene Therapy ? 較少涉及倫理問題， Less ethics problems ? 臨床迫切需要， It is needed urgently in the clinic. ? 腫瘤的基因治療已走在前列。 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(xì)胞因子（干擾素、白介素）的基因?qū)朊庖呒?xì)胞，誘導(dǎo)機(jī)體對(duì)病毒的細(xì)胞和體液免疫應(yīng)答。 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(jì) 80年代發(fā)展起來的最具革命性的醫(yī)療技術(shù)。 美國的基因治療陷入全面低潮。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 確保基因治療技術(shù)真正用于解除患者病痛，造福于人類。 :26:3823:26Feb2327Feb23 1故人江海別，幾度隔山川。 2023年 2月 下午 11時(shí) 26分 :26February 27, 2023 1行動(dòng)出成果，工作出財(cái)富。 23:26:3823:26:3823:262/27/2023 11:26:38 PM 1成功就是日復(fù)一日那一點(diǎn)點(diǎn)小小努力的積累。 下午 11時(shí) 26分 38秒 下午 11時(shí) 26分 23:26: 沒有失敗，只有暫時(shí)停止成功！。 :26:3823:26:38February 27, 2023 1他鄉(xiāng)生白發(fā)，舊國見青山。 , February 27, 2023 雨中黃葉樹，燈下白頭人。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。 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(duì)基因治療產(chǎn)品、方法都采取嚴(yán)格措施監(jiān)控。s disease ) 3/20/2023 147 More than million Americans suffer from Parkinson39。 ? The only choice is to save cancer patients. 3/20/2023 133 治療策略 Strategies of Therapy： ? 導(dǎo)入抑癌基因以抑制癌的發(fā)展、轉(zhuǎn)移 Transfer of tumor suppressor genes to inhibit tumor growth and metastasis. 3/20/2023 134 ? 干擾癌基因的轉(zhuǎn)錄和翻譯，抑制癌基因活性 Interfere with transcription or translation of oncogene in order to inhibit oncogene activity 3/20/2023 135 ? 對(duì)瘤組織進(jìn)行細(xì)胞因子修飾，增強(qiáng)腫瘤細(xì)胞的免疫原性 Modify cytokine gene expression in tumor tissue by transferring genes to enhance immune response. ? 導(dǎo)入自殺基因，殺傷癌細(xì)胞 Transfer of suicide genes to cause death of cancer cells. 3/20/2023 136 三、病毒病的基因治療 Gene Therapy of virus disease 抗病毒基因治療十分引人注目。在以后的 10個(gè)月內(nèi)她又接受了 7次這樣的治療，同時(shí)也接受酶治療。安德森等人對(duì)腺甘脫氨酶缺乏癥（ ADA缺乏癥）的基因治療，是世界上第一個(gè)基因治療成功的范例。 Asialoglycoprotein receptor(ASGP—R) is only express on the surface of hepatocytes, and its natural ligand is asialoglycoprotein. DNA plex carrying the ligand can be transferred directionally into hepatocytes. 3/20/2023 114 (DNA injection) – 直接將裸 DNA注入動(dòng)物或器官組織內(nèi)。 The specific receptors of cell membrane can bine with corresponding ligands ,and transfer the ligands into cells via endocytosis. 3/20/2023 110 3/20/2023 111 ? 將外源 DNA與已證明有細(xì)胞或組織親和性的配體偶聯(lián)，可使 DNA具有靶向性。 Packaging cells are a kind of special cell lines that can enable defective retrovirus (helper virus ) that lack packaging signals and correlation sequence to transfer into mammal cells. 3/20/2023 101 ? 將上兩部分結(jié)合使用即可產(chǎn)生攜帶治療基因，而只有一次感染能力的病毒顆粒 假病毒顆粒（ pseudoviral particles） Combine above two part to form pseudoviral particles that carry therapy gene and can infect (or transduce) target cells only once. 3/20/2023 102 ? 假病毒感染效率非常高 High efficiency of transfection. ? 用于體內(nèi)基因治療時(shí)，只能感染增殖期細(xì)胞，有一定局限性。 A method of letting a foreign functional gene express by transferring the gene into target cells (eg. Lymphocytes, hematopoietic stem cells, skin or