Oregon Health & Science University, Portland, OR, USA.
Blood. 2012 Mar 1;119(9):1992-2002. doi: 10.1182/blood-2011-06-354647. Epub 2012 Jan 10.
Fanconi anemia, complementation group C (FANCC)-deficient hematopoietic stem and progenitor cells are hypersensitive to a variety of inhibitory cytokines, one of which, TNFα, can induce BM failure and clonal evolution in Fancc-deficient mice. FANCC-deficient macrophages are also hypersensitive to TLR activation and produce TNFα in an unrestrained fashion. Reasoning that suppression of inhibitory cytokine production might enhance hematopoiesis, we screened small molecules using TLR agonist-stimulated FANCC- and Fanconi anemia, complementation group A (FANCA)-deficient macrophages containing an NF-κB/AP-1-responsive reporter gene (SEAP). Of the 75 small molecules screened, the p38 MAPK inhibitor BIRB 796 and dasatinib potently suppressed TLR8-dependent expression of the reporter gene. Fanconi anemia (FA) macrophages were hypersensitive to the TLR7/8 activator R848, overproducing SEAP and TNFα in response to all doses of the agonist. Low doses (50nM) of both agents inhibited p38 MAPK-dependent activation of MAPKAPK2 (MK2) and suppressed MK2-dependent TNFα production without substantially influencing TNFα gene transcription. Overproduction of TNFα by primary FA cells was likewise suppressed by these agents and involved inhibition of MK2 activation. Because MK2 is also known to influence production and/or sensitivity to 2 other suppressive factors (MIP-1α and IFNγ) to which FA hematopoietic progenitor cells are uniquely vulnerable, targeting of p38 MAPK in FA hematopoietic cells is a rational objective for preclinical evaluation.
范可尼贫血,补体组 C(FANCC)缺陷的造血干细胞和祖细胞对多种抑制性细胞因子敏感,其中一种细胞因子 TNFα 可诱导 Fancc 缺陷小鼠的 BM 衰竭和克隆进化。FANCC 缺陷的巨噬细胞对 TLR 激活也敏感,并以不受限制的方式产生 TNFα。由于推测抑制抑制性细胞因子的产生可能增强造血作用,我们使用 TLR 激动剂刺激的含有 NF-κB/AP-1 反应性报告基因(SEAP)的 FANCC 和范可尼贫血,补体组 A(FANCA)缺陷巨噬细胞筛选小分子。在筛选的 75 种小分子中,p38 MAPK 抑制剂 BIRB 796 和 dasatinib 强烈抑制 TLR8 依赖性报告基因的表达。FA 巨噬细胞对 TLR7/8 激活剂 R848 高度敏感,对所有剂量的激动剂均过度产生 SEAP 和 TNFα。两种药物的低剂量(50nM)抑制 p38 MAPK 依赖性 MAPKAPK2(MK2)的激活,并抑制 MK2 依赖性 TNFα 产生,而不会对 TNFα 基因转录产生重大影响。这些药物也抑制原发性 FA 细胞中 TNFα 的过度产生,这涉及抑制 MK2 激活。因为 MK2 也已知会影响到 FA 造血祖细胞特有的另外两种抑制性因子(MIP-1α 和 IFNγ)的产生和/或敏感性,因此在 FA 造血细胞中靶向 p38 MAPK 是临床前评估的合理目标。