自噬性组蛋白密码在骨髓增生异常综合征病机转化中的调控机制及中药干预研究

Autophagy is closely related to the pathogenesis of myelodysplastic syndrome (MDS), a clonal disease originating from hematopoietic myeloid stem cells. The accumulation of metabolites caused by defective autophagy plays an important role in the pathogenesis of MDS and its conversion to acute myeloid leukemia (AML). The process of autophagy occurs during the transcription of histones and the post-translational genetic modification of epigenetics. In these processes, autophagy related histone modification, also named "autophagic histone code" has become the main stream. Basing on the previous series of researches on "apoptotic histone code" for MDS and their results, this study is to further explore the effect of histone modification on the autophagy regulation and traditional Chinese medicine (TCM) syndrome transformation in MDS, and to investigate the intervention mechanism of Chinese medicine. In this study, the following hypotheses will be confirmed by using FCM and Western Blot: 1, All different risks stratification patients with MDS have autophagic defects, and there is a difference in the level of autophagy among these patients, which is an important cause for MDS syndrome type of transformation from "right vacuity" to "stasis toxin"; 2. The reduction of autophagy causes ROS accumulation, which further inhibit the autophagy. This may be the molecular mechanism of "stasis toxin causing carcinoma" in TCM. 3. The intervention of spleen-fortifying kidney-supplementing and toxin-resolving Chinese medicine in MDS cell line regulates the autophagy pathway and reduces ROS level through histone modification.

自噬与起源于造血髓系定向干细胞的克隆性疾病——骨髓增生异常综合征（MDS）的发生密切相关，自噬缺陷引起的代谢产物堆积在MDS发病及向急性髓系白血病（AML）转化过程中扮演了重要角色。自噬过程囊括了转录和包括组蛋白翻译后修饰在内的表观遗传程序，后者即“自噬性组蛋白密码”。本项研究基于前期关于MDS的“凋亡性组蛋白密码”系列研究及成果，拟进一步探索MDS发病相关的组蛋白修饰对自噬调控和中医证型转化的影响及中药干预机制，采用流式术、Western Blot等方法证实：1、MDS患者具有自噬缺陷，且不同危度自噬水平存在差异性，是导致MDS“正虚”、“瘀毒”证型转化的重要原因；2、自噬下降引起ROS堆积，ROS升高抑制自噬的恶性循环是中医“瘀毒致癌”的分子机制；3、健脾补肾解毒中药干预MDS细胞株通过组蛋白修饰调控特定自噬通路抑制ROS水平。

背景：自噬与起源于造血髓系定向干细胞的克隆性疾病——骨髓增生异常综合征(MDS)的发生密切相关，自噬缺陷引起的代谢产物堆积在MDS发病及向急性髓系白血病(AML)转化过程中扮演了重要角色。本项研究基于前期关于MDS的“凋亡性组蛋白密码”系列研究及成果，聚焦细胞自噬现象在MDS发病与转化中作用， 进一步探索MDS发病相关的组蛋白修饰对自噬调控和中医证型转化的影响及中药干预机制，为中医药对MDS不同证型及危度BMMNCs的表观遗传学干预机制和辨证论治、预防复发转化提供新的理论依据。.主要研究内容：比较MDS不同证型与正常人BMMNCs自噬水平的差异性，分析MDS不同证型代谢产物ROS水平，探索MDS不同证型H4K16乙酰化修饰与自噬水平的相关性，证明中药复方提取物干预MDS细胞通过改变组蛋白密码调控自噬及ROS水平，并筛选组蛋白修饰调控自噬的主要信号通路。.重要结果及关键数据：MDS患者具有自噬缺陷，正虚毒蕴组和毒盛正虚组患者BMMNCs的自噬水平均低于正常组，且毒盛正虚组BMMNCs的自噬水平显著低于正虚毒蕴组，表明自噬与 MDS 发病和进展具有相关性。正虚毒蕴组和毒盛正虚组代谢产物ROS水平均显著高于正常组，且毒盛正虚组ROS水平更高，表明代谢产物ROS是MDS疾病进展中“瘀毒”病理因素的生物学基础。H4K16乙酰化是MDS疾病进展中的致病因子，MDS患者H4K16乙酰化水平低于正常人，对自噬具有正向调控作用。健脾补肾解毒中药三子黄石散可以诱导MDS细胞自噬及降低有毒代谢产物ROS水平，并通过改变组蛋白密码调控自噬。.科学意义：通过细胞自噬与组蛋白密码修饰来揭示MDS“正虚”、“瘀毒”证型演变的现代医学规律，为MDS“虚毒并治”临床策略提供实验室证据。