Activating PTPN11 mutants promote hematopoietic progenitor cell-cycle progression and survival.
Activating PTPN11 mutants promote hematopoietic progenitor cell-cycle progression and survival.
复制标题
DOI：
10.1016/j.exphem.2008.04.016
复制
发表时间：
2008-10
影响因子：
2.6
通讯作者：
Chan, Rebecca J.
中科院分区：
文献类型：
--
作者：
Yang, Zhenyun;Li, Yiping;Yin, Fuqin;Chan, Rebecca J.
PTPN 11编码蛋白酪氨酸磷酸酶Shp 2，突变通常见于青少年粒单核细胞白血病（JMML）。我们假设PTPN 11突变促进细胞周期进程并增强造血祖细胞的存活。用pMIEG 3、pMIEG 3-WT Shp 2、pMIEG 3-Shp 2D 61 Y或pMIEG 3-Shp 2 E76 K转导小鼠骨髓低密度单核细胞，随后进行细胞周期和存活功能分析以及关键细胞周期和程序性细胞死亡调节蛋白的生化分析。与用空载体（MIEG 3）或WT Shp 2转导的细胞相比，响应于低剂量的GM-CSF，更高比例的携带功能获得性Shp 2突变体的造血祖细胞驻留在细胞周期的S或G2期。同样，与用空载体或WT Shp 2转导的细胞相比，表达Shp 2D 61 Y或Shp 2 E76 K的造血细胞显示基于膜联蛋白V染色的凋亡减少，并且在基本培养基中48小时后产生增加的祖细胞集落。为了区分增强的存活与过度增殖，用PKH 26对细胞进行染色以区分未分裂的细胞与分裂的后代。表达Shp 2D 61 Y或Shp 2 E76 K的PKH 26+细胞同样表现出凋亡减少。生化分析后，Akt和ERK反应性细胞周期和程序性细胞死亡调节蛋白的表达发生改变，包括细胞周期蛋白D1，Bcl 2和Bcl XL水平升高，p27，p21和Bim水平降低。总的来说，这些数据表明，功能获得性Shp 2突变体促进造血祖细胞周期进展和存活，并意味着靶向细胞周期或促进细胞凋亡的药物可能对JMML具有治疗潜力。
Mutations in PTPN11, which encodes the protein tyrosine phosphatase Shp2, are commonly found in juvenile myelomonocytic leukemia (JMML). We hypothesized that PTPN11 mutations promote cell cycle progression and confer enhanced survival to hematopoietic progenitors. Murine bone marrow low density mononuclear cells were transduced with pMIEG3, pMIEG3-WT Shp2, pMIEG3-Shp2D61Y, or pMIEG3-Shp2E76K followed by cell cycle and survival functional analysis as well as biochemical analysis for key cell cycle and programmed cell death regulatory proteins. A higher proportion of hematopoietic progenitors bearing the gain-of-function Shp2 mutants were residing in the S or G2 phase of the cell cycle in response to low doses of GM-CSF compared to cells transduced with empty vector (MIEG3) or with WT Shp2. Likewise, Shp2D61Y-or Shp2E76K-expressing hematopoietic cells demonstrated reduced apoptosis based on annexin V staining and produced increased progenitor colonies after 48 hours in minimal media compared to cells transduced with empty vector or WT Shp2. To differentiate enhanced survival v. hyperproliferation, cells were stained with PKH26 to distinguish undivided cells from divided progeny. Shp2D61Y- or Shp2E76K-expressing PKH26+ cells similarly demonstrated reduced apoptosis. Upon biochemical analysis, expression of Akt- and Erk-responsive cell cycle and programmed cell death regulatory proteins were altered, including increased levels of cyclin D1, Bcl2, and BclXL and reduced levels of p27, p21, and Bim. Collectively, these data demonstrate that gain-of-function Shp2 mutants promote hematopoietic progenitor cell cycle progression and survival and imply that agents targeting the cell cycle or promoting apoptosis may have therapeutic potential in JMML.
登录
查看更多内容
影响因子：
20.3
作者：
Burgess, GS;Williamson, EA;Boswell, HS
通讯作者：
Boswell, HS
影响因子：
5.3
作者：
DOBROWOLSKI, S;HARTER, M;STACEY, DW
通讯作者：
STACEY, DW
影响因子：
8
作者：
Kawada, M;Yamagoe, S;Uehara, Y
通讯作者：
Uehara, Y
影响因子：
5.3
作者：
Kerkhoff, E;Rapp, UR
通讯作者：
Rapp, UR
影响因子：
20.3
作者：
Chan, RJ;Leedy, MB;Kapur, R
通讯作者：
Kapur, R