【作者】 唐勇；

【导师】 王椿；

【作者基本信息】 上海交通大学 ， 内科学（血液）， 2017， 博士

【摘要】 第一部分mir-155靶向SOCS1调控的JAK2-STAT5信号途径在原发性骨髓纤维化发生中的潜在作用背景:络氨酸激酶JAK-STAT信号通路的持续激活,是众多血液肿瘤,尤其是骨髓增值性肿瘤(MPN)的常见病因学事件,但其调控机制尚未完全阐明。许多研究证实micro RNA-155在众多肿瘤中高表达且与肿瘤的发生及进展密切相关。目的:揭示原发性骨髓纤维化中micro RNA-155参与激活络氨酸激酶信号转导通路的机制。方法:Real-time PCR检测micro-RNA及SOCS1基因表达;以逆转录病毒为载体过表达mir-155;MTT测定细胞的存活率;Western blot检测靶基因SOCS1以及下游信号JAK2/STAT5的表达;荧光素酶报告基因检测验证mir-155与SOCS1之间的靶效应。结果:在本研究中,我们发现原发性骨髓纤维化患者骨髓中单个核细胞的micro RNA-155表达显著升高,而SOCS1m RNA表达水平则显著降低;以Ba F3作为细胞模型,过表达micro RNA-155可通过上调细胞JAK2-STAT5信号,增加细胞对IL-3的敏感性。进一步实验证实,micro RNA-155通过调节靶基因SOCS1的表达进而影响JAK2-STAT5信号途径发挥上述生物学效应。结论:mir-155是通过调节靶基因SOCS1的表达进而影响JAK2/STAT5信号途径发挥对BAF3细胞对IL-3敏感性的调节。上述结果有助于为我们理解PMF的发病机制提供新的视角。第二部分以每周一次1.6mg/m2万珂为基础的BCD方案对初发老年不适合标准剂量化疗的多发性骨髓瘤患者的疗效评价背景:硼替佐米联合烷化剂有抗骨髓瘤的效应。但是由于其神经毒性,在临床应用中受到限制。对于老年初发的多发性骨髓瘤患者,有些不适合标准剂量治疗。而使用1.6mg/m2万珂联合环磷酰胺和地塞米松的BCD方案在这一领域并无研究。目的:我们报道了对这部分患者应用静脉内1.6mg/m2万珂为基础的BCD方案的经验。评估其有效性及安全性。方法:在2010年3月开始至末次随访至2015年2月,总共有34例初发的老年患者接受了BCD方案(万珂1.6mg/m2静脉注射d1/8,环磷酰胺200mg/m2静脉滴注d1-4,地塞米松20mg静脉滴注d1-4,8-11)。结果:34例患者中,总共有14例(41%)患者获得完全缓解,6例(18%)患者获得非常好的部分缓解,10例(29%)获得部分缓解。总有效率达88%。治疗2个疗程后,获得非常好的部分缓解和完全缓解的患者与较获得部分缓解及耐药的患者相比,无进展生存时间(21.4个月vs10.6个月,p=0.002)和总生存时间(23.0 vs.16.8个月,p=0.043)均明显延长。2年的无进展生存率及总生存率分别达到26.5和64.7%。发生Ⅰ/Ⅱ级神经毒性见于20%的化疗次数,并无观察到化疗中出现Ⅲ/Ⅳ级神经毒性。没有患者因为神经毒性或其它不良反应退组。结论:以1.6mg/m2万珂为基础的BCD方案对于老年初发不适合标准剂量的多发性骨髓瘤患者是有效而安全的方案。第三部分培门冬酶+GOD方案一线治疗进展期结外NK/T细胞淋巴瘤的回顾性分析背景:进展期结外NK/T细胞淋巴瘤预后极差,由于临床侵袭性高,联合化疗效果差。不受多药耐药的联合化疗是目前进展期NK/T细胞淋巴瘤的推荐治疗方案。目的:本研究回顾性分析了培门冬酶+GOD方案一线治疗进展期NK/T细胞淋巴瘤的有效性和安全性,同时也收集和分析了临床预后因素。方法:2011年1月至2014年12月,共有18例Ⅲ/Ⅳ期NT/T细胞淋巴瘤患者入组。所有患者接受3~6个疗程Pegasp-GOD方案化疗。结果:3疗程后ORR67%(12/18),CR率28%(5/18),PR率39%(7/18)。平均随访时间18个月(3~24个月)。平均OS10个月,PFS8.5个月。疗效为PR及以上患者OS及PFS较无效者显著延长(平均OS分别为15个月、10个月,P=0.001;平均PFS分别为15个月和7个月,P=0.001)。同时,诱导治疗结束后血浆EBV-DNA水平更低的患者OS及PFS时间显著延长。结论:该方案不良反应可以耐受,是治疗该病有效且低毒的方案,值得进一步开展前瞻性研究。更多还原

【Abstract】 Part 1 The potential role of targeting the SOCS1-regulated JAK2-STAT5 signaling pathway by mi R-155 in the development of primary myelofibrosisBackground: Sustained activation of the Janus kinase(JAK)-signal transducer and activator of transcription(STAT)signaling pathway is a common etiological event in a variety of hematological tumors,especially myeloproliferative neoplasms(MPNs).However,the regulatory mechanism has not been fully elucidated.Numerous studies have demonstrated that micro RNA-155(mi R-155)is highly expressed in many tumors and is closely related to tumor development and progression.Objective: The purpose of the present study was to investigate whether mi R-155 participated in the pathogenesis of primary myelofibrosis(PMF)through regulation of the JAK2-STAT5 signaling pathway.Methods: Real-time polymerase chain reaction(PCR)was performed to examine micro RNA and the suppressor of cytokine signaling 1(SOCS1)gene expression.A retrovirus was used as a mi R-155 overexpression vector.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was performed to quantify the cell survival rate.Western blotting was performed to analyze the expression of SOCS1 and its downstream signaling molecules JAK2/STAT5.The luciferase reporter gene assay was employed to verify the target effect of mi R-155 on SOCS1.Results: In the present study,compared to healthy individuals,the mi R-155 expression level was significantly increased and the SOCS1 m RNA and protein expression levels were markedly reduced in bone marrow mononuclear cells collected from PMF patients.Using Ba F3 as a cell model,we discovered that mi R-155 overexpression enhanced the sensitivity of the cells to interleukin-3(IL-3)by upregulating JAK2-STAT5 signaling.Further experiments demonstrated that mi R-155 affected the JAK2-STAT5 signaling pathway by regulating the expression of its target gene SOCS1 to exert the biological effects described above.Conclusion: mi R-155 affects JAK2/STAT5 signaling by regulating the expression of its target gene SOCS1,thereby exerting a regulatory effect on the sensitivity of Ba F3 cells to IL-3.The above results provide a new perspective for the pathogenic mechanisms of PMF.Part 2 Once-weekly 1.6mg/m2 bortezomib BCD regimen in elderly patients with newly diagnosed multiple myeloma who are unfit for standard dose chemotherapyBackground: Bortezomib has shown anti-myeloma effects in combination with alkylating agents,but clinical benefits can be limited by neurotoxicity.There is less information on the efficacy and tolerability of once-weekly 1.6mg/m~2 bortezomib BCD regimen in elderly patients with newly diagnosed MM who are unfit for standard dose chemotherapy Objective: Here,we report our experience of weekly 1.6mg/m~2 intravenous bortezomib in this group of patients,access the efficacy and safety of this program。Methods: Between March 2010 and February 2015,we treated 34 newly diagnosed elderly patients with the combination of bortezomib 1.6 mg/m~2 intravenously on days 1 and 8;cyclophosphamide 200mg/m~2 intravenously on days 1–4;dexamethasone 20 mg intravenously on days 1-4,and 8-11.Results: Among the 34 patients,14(41%)responded with CR,6(18%)with VGPR and 10(29%)with PR.The ORRs were 88%.After 2 cycles of treatments,the survival of patients who attained a response of VGPR or CR was significantly longer than those with PR or resistance to BCD,for both progression-free survival(PFS)(21.4 vs.10.6 mo,p=0.002)and overall survival(OS)(23.0 vs.16.8 mo,p=0.043).The 2-year PFS and OS were 26.5% and 64.7% respectively in these elderly MM patients in our study.Grade 1/2 neuropathy was observed in 20% of the cycles while grade 3/4 neuropathy was not observed.No patients withdrew due to neuropathy or other side effects.Conclusion: Once-weekly bortezomib at 1.6mg/m2 BCD regimen is both effective and safe in elderly patients with newly diagnosed MM who are unfit for standard dose chemotherapy.Part 3 Retrospective study of Pegaspargase,Gemicitabine,oxaliplatin and dexamethasone(Peg-Gem OD)as a first-line therapy for advanced-stage extranodal NK/T cell lymphomaBackground: The prognosis of advanced-stage extranodal NK/T cell lymphoma(ENKTL)is extremely poor because of its very aggressive clinical course and unsatisfactory response to conventional combination chemotherapy.Systemic chemotherapy containing drugs unaffected by multidrug resistance pathway are currently recommended for patients with advanced-stage ENKTL.Objective: This study was conducted to retrospectively investigate the efficacy and safety of pegaspargase,gemicitabine,oxaliplatin and dexamethasone(Peg-Gem OD)combination chemotherapy as a first-line therapy for advanced-stage ENKTL.Methods: Clinical prognostic factors from these patients were also collected and analyzed.A cohort of 18 patients with newly diagnosed stage III/IV ENKTL who received Peg-Gem OD regimen as a first-line treatment was retrospectively identified from January 2011 to December 2014.Results: All patients were subjected to 3 to 6 cycles of Peg-Gem OD chemotherapy.After 3 cycles of therapy,the overall response rate(ORR)was 67%(12/18)with a complete response(CR)rate of 28%(5/18)and a partial response(PR)rate of 39%(7/18).The median follow-up period was 18 months(range 3~24 months).The median overall survival(OS)and progression-free survival(PFS)time were 10 and 8.5 months respectively.For those responders(CR+PR),the median OS and PFS time were significantly better than those of non-responders(median OS,15 months vs.10 months;P=0.001 and median PFS,15 months vs.7 months;P=0.001).Furthermore,patients with low plasma EBV-DNA levels after induction chemotherapy had a remarkably longer OS and PFS time(median OS,18 months and median PFS,18 months).The toxicity of Peg-Gem OD regimen was acceptable.Conclusion: The Peg-Gem OD regimen is highly effective and tolerable first-line chemotherapy for newly diagnosed advanced-stage ENKTL and is worth further study in prospective trial.更多还原