Limiting numbers of G156A O6-methylguanine–DNA methyltransferase-transduced marrow progenitors repopulate nonmyeloablated mice after drug selection
Limiting numbers of G156A O6-methylguanine–DNA methyltransferase-transduced marrow progenitors repopulate nonmyeloablated mice after drug selection
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药物选择后，有限数量的 G156A O6-甲基鸟嘌呤-DNA 甲基转移酶转导的骨髓祖细胞重新填充非清髓小鼠
DOI：
10.1182/blood.v95.10.3078
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发表时间：
2000
期刊：
影响因子：
20.3
通讯作者：
S. Gerson
中科院分区：
文献类型：
--
作者：
B. M. Davis;O. Koç;S. Gerson
造血基因治疗的有限疗效可以通过体内选择转导的长期再填充细胞(LTRC)来改善。我们选择G156A mGMT(▵mGMT)转导的LTRC存在于5 × 10 4到10 0 × 10 4的骨髓细胞中，每隔3~4周给非去髓小鼠输注O6-苄基鸟嘌呤(BG)和BCNU。为了促进植入，小鼠在输液前给予非消融性剂量的BG和BCNU。未经筛选，输注后24~30周，所有造血祖细胞集落形成单位均未检测到▵-mGMT。BG、BCNU后，▵、MGMT+CFU出现频率较高，其比例随治疗周期的延长而增加。在BG和BCNU的2-3个周期后，许多小鼠稳定地重建了75%到100%的▵-MGMT+CFU至少6个月，相当于940倍的浓缩。因此，BG和BCNU的干细胞毒性使▵MGMT转导的LTRC能够重新填充骨髓。在非去髓小鼠中，这种程度的选择压力远远大于之前的耐药基因转移研究中观察到的压力。这些数据支持我们批准的临床试验，选择耐药的、转导的造血细胞，潜在地减少癌症患者药物诱导的累积骨髓抑制。这些数据还表明，▵MGMT可能是一种有效的、显性的、可选择的标记，用于双基因治疗。
The limited efficacy of hematopoietic gene therapy can be improved by in vivo selection for transduced long-term repopulating cells (LTRC). We selected for G156A MGMT (▵MGMT) transduced LTRC present in 5 × 104 to 100 × 104 marrow cells infused into nonmyeloablated mice by the administration of O6-benzylguanine (BG) and BCNU every 3 to 4 weeks. To facilitate engraftment, mice were given a nonablative dose of BG and BCNU before infusion. Without selection, ▵MGMT was not detected in any hematopoietic colony-forming units (CFU) 24 to 30 weeks after infusion. After BG and BCNU, ▵MGMT+ CFU were frequently detected, and their proportions increased with each treatment cycle. After 2 to 3 cycles of BG and BCNU, many mice were stably reconstituted with 75% to 100% ▵MGMT+ CFU for at least 6 months, representing up to 940-fold enrichment. Thus, BG and BCNU stem cell toxicity allows ▵MGMT-transduced LTRC to repopulate the bone marrow. This degree of selection pressure in nonmyeloablated mice is far greater than that observed in previous drug-resistance gene transfer studies. These data support our approved clinical trial to select for drug-resistant, transduced hematopoietic cells, potentially decreasing cumulative drug-induced myelosuppression in patients with cancer. These data also suggest that ▵MGMT may be a potent, dominant, selectable marker for use in dual gene therapy.
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影响因子：
11.2
作者：
Prescilla E. Gonzaga;Philip M. Potter;T. Niu;Dong Yu;D. Ludlum;Joseph A Rafferty;G. P. Margison;T. Brent
通讯作者：
Prescilla E. Gonzaga;Philip M. Potter;T. Niu;Dong Yu;D. Ludlum;Joseph A Rafferty;G. P. Margison;T. Brent
人类 CD34 造血祖细胞具有低水平、细胞因子无反应的 O6-烷基鸟嘌呤-DNA 烷基转移酶，并且对 O6-苄基鸟嘌呤加 BCNU 敏感。
影响因子：
20.3
作者：
Gerson,SL;Phillips,W;Kastan,M;Dumenco,LL;Donovan,C
通讯作者：
Donovan,C
通过对移植小鼠进行 BCNU 处理，人烷基转移酶转导的小鼠骨髓祖细胞在体内得到富集。
DOI：
--
发表时间：
1997
期刊：
Experimental hematology.
影响因子：
--
作者：
Allay,JA;Davis,BM;Gerson,SL
通讯作者：
Gerson,SL
影响因子：
2.9
作者：
PEGG, AE;BOOSALIS, M;DOLAN, ME
通讯作者：
DOLAN, ME
影响因子：
56.9
作者：
DUMENCO, LL;ALLAY, E;GERSON, SL
通讯作者：
GERSON, SL