肿瘤的造血干细胞多药耐药基因治疗研究进展

造血干细胞具有多系分化潜能及自我更新能力，其生命周期长，是基因转移的理想靶细胞。这种转基因的造血干细胞可在造血组织中长期稳定表达目的产物，而较成熟的祖细胞作为靶细胞时其基因表达相对短暂。将多药耐药（ＭＤＲ１）基因转移至造血干细胞，在体内存在抗癌药物时具有选择优势，可提高化疗剂量，保护骨髓不受高剂量化疗药物损伤。本文从以下几个方面综述造血干细胞ＭＤＲ基因疗法的原理、临床应用方案及其前景。     

IL-6基因疗法增强同基因骨髓移植后的造血重建

目前临床上治疗无法手术的恶性肿瘤仍以放化疗为主，虽然增加放化疗剂量可显著提高对肿瘤的治疗效果．但在增加放化疗剂量的同时可导致机体造血功能严重低下，重者则导致致死性骨髓抑制，因此这使降低放化疗剂量或中止放化疗，这样往往影响肿瘤的治疗效果。研究表明细胞因子如IL-3、IL-6、GM-CSF等能显著地促进放化疗后机体造血功能的尽早恢复，     

异体外周血造血干细胞移植治疗恶性肿瘤

继骨髓移植，脐血移植和自体外周造血干细胞移植之后，最后异体外周血造血干细胞移植的成功，为临床治疗恶性肿瘤及血液病开辟了新的有效途径。现有资料表明，经造血因子动员的供体外周血造血干／细胞含量及增殖能力的骨髓相当，异体樾入迅速，能够支持肿瘤病人长期造血。     

自体造血干细胞移植后的免疫重建研究进展

大剂量放化疗联合造血干细胞移植是治疗恶性血液肿瘤及部分实体瘤的新疗法之一。在世界范围内,每年大约有1万例自体造血干细胞移植用来治疗恶性肿瘤。近年来的各项随机实验证实,大剂量化疗联合自体造血干细胞输注可明显延长血液系统恶性疾病包括复发的非霍奇金淋巴瘤犤1犦,急性髓性白血病犤2犦和多发性骨髓瘤犤3犦患者的无病生存期(DFS)。在实体瘤的治疗上也已取得了较好的效果犤4犦。然而,移植后仍有部分病人出现复发和转移,移植后免疫功能的长期抑制是造成移植后恶性肿瘤复发的重要原因之一,移植后免疫功能的状态及免疫治疗对清除体内残…     

细胞因子对化疗损伤的造血调控

细胞因子对造血细胞的生存、增殖和分化起重要的调控作用,化疗可引起造血衰竭及骨髓抑制,而正、负性造血因子在促进化疗后造血恢复方面有不同的作用和机制.本文概述了有关细胞因子对化疗后造血恢复的调控作用及其机制,为基础和临床研究提供了理论依据,也为开发和研制新的化学保护剂提供了思路.     

造血干细胞移植在儿童白血病治疗中的应用

 白血病是儿童时期最常见的恶性肿瘤,约占该时期所有恶性肿瘤的35 %,对于传统化疗难以治愈的高危急性白血病和慢性粒细胞白血病,造血干细胞移植（HSCT）是主要的治疗手段。目前国际上推荐首选HLA匹配相关供者（MRD）的HSCT。由于80 %的儿童白血病患者缺少这种供者,自体造血干细胞移植（auto-HSCT）也被选择性地应用于急性淋巴细胞白血病（ALL）和急性髓系白血病（AML）的治疗。据统计有30 %～40 %的患者接受了匹配的无关供者（MUD）的HSCT治疗。近年来,无关脐血移植（UDCBT）和HLA不相匹配的相关供者HSCT呈上升趋势。就近年来该方面的进展作一综述。     

细胞因子对化疗损伤的造血调控

细胞因子对造血细胞的生存、增殖和分化起重要的调控作用，化疗可引起造血衰竭及骨髓抑制，而正、负性造血因子在促进化疗后造血恢复方面有不同的作用和机制。本文概述了有关细胞因子对化疗后造血恢复的调控作用及其机制，为基础和临床研究提供了理论依据，也为开发和研制新的化学保护剂提供了思路。     

新药时代造血干细胞移植在多发性骨髓瘤治疗中的地位

 与传统化疗相比,自体造血干细胞移植可提高多发性骨髓瘤（MM）患者的缓解率,延长无进展生存期,是治疗MM的一线方案。但近年来,基于新型药物的联合诱导、巩固和维持治疗提高了MM的治疗效果,对自体造血干细胞移植的地位构成了挑战。异基因造血干细胞移植虽然具有治愈MM的潜能,但移植相关死亡率高,患者的总体生存并未获益。而减低剂量预处理异基因移植虽降低了移植相关死亡率,具有一定的移植物抗骨髓瘤作用,但移植物抗宿主病的发生率高。文章总结了MM干细胞移植相关的临床试验结果,旨在定义新药时代造血干细胞移植在MM治疗中的地位。     

活化的造血—免疫干细胞移植

对化疗敏感的肿瘤，加大化疗药物的强度可以提高治疗效果。PMT、PB8CT能够明确改善高剂量化疗的相关并发症，为了缩短骨髓再生低下期，减少感染，减少移植物中的肿瘤细胞污染，利用体外筛选和培养活化的造血-免疫干、祖细胞移植成为发展的方向。     

造血生长因子在肿瘤化疗中应用的研究进展

众多特异的造血生长因子调控着人体造血,给肿瘤患者的化、放疗提供了有力的支持.重组人粒细胞集落刺激因子(rhG-CSF)的应用减少了化疗导致的持续而严重的中性粒细胞减少症和相关感染的发生,使外周血干细胞动员高效安全,加速干细胞移植后的造血功能重建.聚乙二醇重组人粒细胞集落刺激因子(PEG-rhG-CSF)半衰期长,每周期1次给药可有效防治发热性中性粒细胞减少引起的感染和非血液系统恶性肿瘤化疗导致的中性粒细胞减少症.干细胞因子在临床中单独使用,并无体外实验的动员效果,只能与rhG-CSF联合使用.防治化疗引起的口腔粘膜已经成为粒/巨噬细胞集落刺激因子的一种新的临床用途.白细胞介素-11是目前惟一被美国FDA批准上市的防治血小板减少的药物.血小板生长因子由于临床效果不确切,并可导致中和抗体的产生,已停止临床研究.促红细胞生成素被广泛用于治疗慢性肾衰引起的贫血,但在肿瘤患者贫血中的应用仍缺乏足够的研究.     

盆腔肿瘤IMRT下骨髓剂量-体积参数与急性血液学毒性相关性研究进展

盆腔恶性肿瘤同期放化疗期间发生骨髓抑制的风险较高,盆腔受照射骨髓(骼)的剂量-体积关系与放化疗期间的急性血液学毒性存在相关,但缺乏此方面公认的参数。盆腔不同部位骨髓(骼)造血能力具有异质性,靠近体中轴的盆腔骨髓造血能力最强,即功能性骨髓。找准盆腔功能性骨髓的剂量-体积参数与盆腔恶性肿瘤放化疗期间急性血液学毒性的关系也许是今后发展方向。     

软组织肉瘤化疗耐药机制及治疗的研究进展

骨与软组织肿瘤发病率低,属于罕见肿瘤范畴,目前化疗仍是骨与软组织肉瘤有效的治疗方法,但化疗耐药的存在严重影响了患者的近期疗效、远期生存,本文对软组织肉瘤化疗耐药机制及相应的治疗进展作一综述。     

Hematopoietic stem cell compartment: Acute and late effects of radiation therapy and chemotherapy

The bone marrow is an important dose-limiting cell renewal tissue for chemotherapy, wide-field irradiation, and autologous bone marrow transplantion. Over the past 5–10 years a great deal has been discovered about the hematopoietic stem cell compartment. Although the toxicity associated with prolonged myelosuppression continues to limit the wider use of chemotherapy and irradiation, ways are being discovered to circumvent this toxicity such as with the increasing use of cytokines. This review describes what is known of how chemotherapy and irradiation damage stem cells and the microenvironment, how cytokines protect hematopoietic cells from radiation damage and speed marrow recovery after chemotherapy or marrow transplantation, and how various types of blood marrow cells contribute to engraftment and long-term hematopoiesis after high doses of cytotoxic agents and/or total body irradiation.     

Adenovirus as a gene therapy vector for hematopoietic cells

Adenovirus (Adv)-mediated gene transfer has recently gained new attention as a means to deliver genes for hematopoietic stem cell (HSC) or progenitor cell gene therapy. In the past, HSCs have been regarded as poor Adv targets, mainly because they lack the specific Adv receptors required for efficient and productive Adv infection. In addition, the nonintegrating nature of Adv has prevented its application to HSC and bone marrow transduction protocols where long-term expression is required. There is even controversy as to whether Adv can infect hematopoietic cells at all. In fact, the ability of Adv to infect epithelium-based targets and its inability to effectively transfect HSCs have been used in the development of eradication schemes that use Adv to preferentially infect and "purge" tumor cell-contaminating HSC grafts. However, there are data supporting the existence of productive Adv infections into HSCs. Such protocols involve the application of cytokine mixtures, high multiplicities of infection, long incubation periods, and more recently, immunological and genetic modifications to Adv itself to enable it to efficiently transfer genes into HSCs. This is a rapidly growing field, both in terms of techniques and applications. This review examines the two sides of the Adv/CD34 controversy as well as the current developments in this field.     

The effect of tumor necrosis factor on normal human hematopoietic progenitors

Tumor necrosis factor-alpha (TNF-alpha), a product of activated macrophages that is cytotoxic to tumor cells, could be used to purge tumor cells from bone marrow before autologous bone marrow transplantation for hematologic malignancies and/or solid tumors. To determine whether exposure to TNF-alpha would have an inhibitory effect on hematopoietic progenitors, we incubated normal human bone marrow with a wide range of concentrations of recombinant human TNF. In order to mimic the conditions that would be used in bone marrow purging, bone marrow cell suspensions were incubated with TNF in doses ranging from 500 to 100,000 U/ml for 24 hours, and were assayed for colony formation in agar. We noted a dose-dependent inhibition of total colony-forming units (CFU) at days 7 and 14, with 50% inhibition occurring at 60,000 U/ml of TNF. TNF exerted a differential effect on CFU so that colony formation by erythroid (CFU-E), multipotential (CFU-GEMM), and macrophage (CFU-M) progenitors was suppressed to a greater extent than that by granulocyte progenitors (CFU-G). However, even after preincubation with TNF at high doses such as 100,000 U/ml, the inhibitory effects of TNF could be abolished by washing cells before culturing. This study demonstrates that hematopoietic precursors survive treatment with TNF at doses that have been shown to be cytotoxic to tumor cells. Although TNF has a significant inhibitory effect on the growth of erythroid, multipotential, and macrophage progenitors in vitro, this effect depends on continuous exposure to TNF for more than 24 hours. Thus, TNF may be useful as a bone marrow purging agent against tumor cells, with relative sparing of normal marrow elements.     

Comparison of different busulfan analogues for depletion of hematopoietic stem cells and promotion of donor-type chimerism in murine bone marrow transplant recipients

Busulfan (1,4-butanediol dimethanesulfonate, BU) is relatively unique among other standard chemotherapy compounds in its ability to deplete noncycling primitive stem cells in the host and consequently to allow for high levels of long-term, donor-type engraftment after bone marrow transplantation (BMT). Such a property explains why this drug can be used as an alternative to total body irradiation in preparative regimes for BMT. However, as with radiation, BU conditioning is still troubled by severe toxicities that limit its applications to suboptimal drug doses. These problems stress the need for other BMT-conditioning drugs that are better tolerated and more selectively targeted toward normal and malignant hematopoietic stem cells. We have therefore compared the effects of various novel dimethanesulfonate compounds (related to BU) in terms of their toxicity to different stem cell subsets in vivo and in vitro and their ability to provide for long-term donor bone marrow engraftment using the congenic glucose-6-phosphate isomerase type 1 marker. Introduction of a benzene or cyclohexane ring in some of these drugs affords rigidity to the molecule and restricts the spatial positioning of the alkylating groups. Among 25 different compounds thus far tested at single doses, PL63 [cis-1,2-(2-hydroxyethyl) cyclohexane dimethanesulfonate] proved to be the most effective in providing for hematopoietic engraftment. The transisomer of the same compound gave significantly less engraftment and was comparable with the effects of dimethylbusulfan and Hepsulfam. The engraftment data correlated well with the depletion of different bone marrow stem cell subsets in the host as measured using the cobblestone area forming cell assay. The extent of stem cell depletion could not be explained on the basis of the distance and orientation of the two alkylating groups. Pharmacokinetic data, however, indicate that there is a correlation between biological activity and plasma levels reached. The diverse cytotoxic effects shown by these novel analogues of BU have provided a basis for relating biological activity with pharmacokinetic properties rather than with structural properties such as distance and orientation of the two alkylating groups. The identification of highly active compounds such as PL63 offers an opportunity for further developing other closely related drugs for potential application in clinical BMT conditioning therapy.     

Fhit-deficient hematopoietic stem cells survive hydroquinone exposure carrying precancerous changes

The fragile FHIT gene is among the first targets of DNA damage in preneoplastic lesions, and recent studies have shown that Fhit protein is involved in surveillance of genome integrity and checkpoint response after genotoxin exposure. We now find that Fhit-deficient hematopoietic cells, exposed to the genotoxin hydroquinone, are resistant to the suppression of stem cell in vitro colony formation observed with wild-type (Wt) hematopoietic cells. In vivo-transplanted, hydroquinone-exposed, Fhit-deficient bone marrow cells also escaped the bone marrow suppression exhibited by Wt-transplanted bone marrow. Comparative immunohistochemical analyses of bone marrow transplants showed relative absence of Bax in Fhit-deficient bone marrow, suggesting insensitivity to apoptosis; assessment of DNA damage showed that occurrence of the oxidized base 8-hydroxyguanosine, a marker of DNA damage, was also reduced in Fhit-deficient bone marrow, as was production of intracellular reactive oxygen species. Treatment with the antioxidant N-acetyl-l-cysteine relieved hydroquinone-induced suppression of colony formation by Wt hematopoietic cells, suggesting that the decreased oxidative damage to Fhit-deficient cells, relative to Wt hematopoietic cells, accounts for the survival advantage of Fhit-deficient bone marrow. Homology-dependent recombination repair predominated in Fhit-deficient cells, although not error-free repair, as indicated by a higher incidence of 6-thioguanine-resistant colonies. Tissues of hydroquinone-exposed Fhit-deficient bone marrow-transplanted mice exhibited preneoplastic alterations, including accumulation of histone H2AX-positive DNA damage. The results indicate that reduced oxidative stress, coupled with efficient but not error-free DNA damage repair, allows unscheduled long-term survival of genotoxin-exposed Fhit-deficient hematopoietic stem cells carrying deleterious mutations.     

Biological purging of breast cancer cells using an attenuated replication-competent herpes simplex virus in human hematopoietic stem cell transplantation

Autologous hematopoietic stem cell transplantation after myelosuppressive chemotherapy is used for the treatment of high-risk breast cancer and other solid tumors. However, contamination of the autologous graft with tumor cells may adversely affect outcomes. Human hematopoietic bone marrow cells are resistant to herpes simplex virus type 1 (HSV-1) replication, whereas human breast cancer cells are sensitive to HSV-1 cytotoxicity. Therefore, we examined the utility of G207, a safe replication-competent multimutated HSV-1 vector, as a biological purging agent for breast cancer in the setting of stem cell transplantation. G207 infection of human bone marrow cells had no effect on the proportion or clonogenic capacity of CD34+ cells but did enhance the proliferation of bone marrow cells in culture and the proportion of CD14+ and CD38+ cells. On the other hand, G207 at a multiplicity of infection of 0.1 was able to purge bone marrow of contaminating human breast cancer cells. Because G207 also stimulates the proliferation of human hematopoietic cells, it overcomes a limitation of other purging methods that result in delayed reconstitution of hematopoiesis. The efficient infection of human bone marrow cells in the absence of detected toxicity suggests that HSV vectors may also prove useful for gene therapy to hematopoietic progenitor cells.     

自体骨髓移植联合MHC单倍体相合淋巴细胞治疗急性髓性白血病的临床疗效

目的:自体骨髓移植联合MHC单倍体相合淋巴细胞治疗急性髓性白血病的治疗效果和安全性。方法:以40例急性髓性白血病患者作为研究对象,随机分为两组各20例,研究组患者使用自体骨髓移植联合MHC单倍体相合淋巴细胞进行治疗,对照组单用自体骨髓移植进行治疗。结果:两组患者的造血系统都得到重建,重建时间以及并发症的发生均没有显著差异。而研究组患者复发率显著降低,且复发时间明显地长于对照组患者。研究组患者3年累积无病生存率明显地高于对照组患者,且差异具有统计学意义。结论:自体骨髓移植联合MHC单倍体相合淋巴细胞治疗能够有效控制急性髓性白血病病情,降低其复发,延长患者生存期。     

单倍型异基因造血干细胞移植治疗难治急性型成人T细胞白血病-淋巴瘤1例并文献复习北大核心

目的:探讨成人T细胞白血病-淋巴瘤(adult T cell leukemia-lymphoma,ATL)的临床特征及治疗。方法:回顾分析我院1例ATL的临床病理特征、治疗转归,并进行相关文献复习。结果:患者女性,44岁,祖籍福建,以皮疹、腹胀及水肿为首发表现,影像学检查提示全身多发代谢增高肿大淋巴结、脾大,腋窝淋巴结、腹壁皮肤活检提示外周T细胞淋巴瘤;经预治疗及多次化疗后效果欠佳,行HTLV I-RNA检测为阳性,诊断ATL急性型明确,后选择其子作为供者行单倍型异基因造血干细胞移植,移植后随访15个月至2022年04月仍存活。结论:由于北方地区HTLV-1感染率较低,遇到化疗效果欠佳的外周T细胞淋巴瘤时需考虑到ATL可能,尽早行相关病毒学检测以尽快明确诊断。单倍型异基因造血干细胞移植可以为ATL患者改善长期预后带来希望。