Lysine-specific demethylase 1 inhibitor rescues the osteogenic ability of mesenchymal stem cells under osteoporotic conditions by modulating H3K4 methylation

Bone tissue engineering may be hindered by underlying osteoporosis because of a decreased osteogenic ability of autologous seed cells and an unfavorably changed microenvironment in these patients. Epigenetic regulation plays an important role in the developmental origins of osteoporosis; however, few studies have investigated the potential of epigenetic therapy to improve or rescue the osteogenic ability of bone marrow mesenchymal stem cells(BMMSCs) under osteoporotic conditions. Here, we investigated pargyline, an inhibitor of lysine-specific demethylase 1(LSD1), which mainly catalyzes the demethylation of the di- and mono-methylation of H3K4. We demonstrated that 1.5 mmol·Lpargyline was the optimal concentration for the osteogenic differentiation of human BMMSCs. Pargyline rescued the osteogenic differentiation ability of mouse BMMSCs under osteoporotic conditions by enhancing the dimethylation level of H3K4 at the promoter regions of osteogenesis-related genes. Moreover, pargyline partially rescued or prevented the osteoporotic conditions in aged or ovariectomized mouse models, respectively. By introducing the concept of epigenetic therapy into the field of osteoporosis, this study demonstrated that LSD1 inhibitors could improve the clinical practice of MSC-based bone tissue engineering and proposes their novel use to treat osteoporosis.

Lysine-specific Demethylase 1 Represses THP-1 Monocyte-to-macrophage Differentiation

Objective To investigate the role of lysine-specific demethylase 1 （LSD1） in the process of THP-1 monocyte-to-macrophage differentiation. Methods Quantitative reverse transcription-polymerase chain reaction （qRT-PCR） and Western blotting were performed to analyze the expression of LSD1 and interleukin-6 （IL-6） in THP-1 monocytes and THP-l-derived macrophages. Chromatin immunoprecipitation （ChiP） assay was applied to detect the occupancy of LSD1 and H3K4 methylation at IL-6 promoter during THP-1 monocyte-to-macrophage differentiation. IL-6 mRNA level and H3K4 methylation at IL-6 promoter were analyzed using qRT-PCR and ChiP assay in LSD 1 -knockdown THP- 1 cells treated with 12-O-tetradecanoylphorbol- 13-acetate （TPA） for 0 4, 8, 12, and 24 hours. Fluorescence activated flow cytometry was performed to reveal the percentage of macrophages differentiated from THP- 1 monocytes. Results The expression of LSD1 reduced during THP-1 monocyte-to-macrophage differentiation （P〈0.01）. LSD1 occupancy decreased and H3K4 methylation increased at IL-6 promoter during the differentiation. With knockdown of LSD1, H3K4 methylation at IL-6 promoter was found increased after TPA treatment at different times points （all P〈0.05, except 24 hours）. The percentage of macrophages increased significantly in theTHP-I cells with LSD1 knockdown （P〈0.05）. Conclusions LSD1 is repressed during the monocyte-to-macrophage differentiation of THP-1 cells. Suppression of LSD 1-mediated H3K4 demethylation may be required for THP-1 monocyte-to-macrophage differentiation.

卵巢癌组织中LSD1、NDRG1基因表达量与癌细胞迁移、侵袭的相关性研究

目的:研究卵巢癌组织中赖氨酸特异性组蛋白去甲基化酶1(lysine-specific demethylase 1,LSD1)、N-myc下游调节基因1(N-myc downstream regulated 1,NDRG1)基因表达量与癌细胞迁移、侵袭的相关性。方法:选择2014年3月~2017年7月扶风县人民医院接受手术切除的卵巢癌患者作为研究对象,手术切除后取卵巢癌组织和癌旁组织,测定LSD1、NDRG1基因及迁移基因、侵袭基因的表达量。结果:卵巢癌组织中LSD1、YKL40、COX2、Twist、IFITM1、CatL、CTHRC1、MMP2、FUNDC1基因的mRNA表达量显著高于癌旁组织,NDRG1、E-cadherin、Wnt5a基因的mRNA表达量显著低于癌旁组织;LSD1高表达的卵巢癌组织中YKL40、COX2、Twist、IFITM1、CatL、CTHRC1、MMP2、FUNDC1的mRNA表达量显著高于LSD1低表达的卵巢癌组织,E-cadherin、Wnt5a基因的mRNA表达量显著低于LSD1低表达的卵巢癌组织。结论:卵巢癌组织中LSD1高表达、NDRG1低表达能够促进癌细胞的迁移、侵袭。

Correlation of LSD1 and NDRG1 gene expression in ovarian cancer tissue with cancer cell migration and invasion

Objective: To study the correlation of LSD1 and NDRG1 gene expression in ovarian cancer tissue with cancer cell migration and invasion. Methods: Patients with ovarian cancer who underwent surgical resection in Fufeng County People's Hospital between March 2014 and July 2017 were selected as the research subjects, and the ovarian cancer tissue and adjacent tissue were collected after surgical resection to determine the expression of LSD1, NDRG1, migration genes and invasion genes. Results: LSD1, YKL40, COX2, Twist, IFITM1, CatL, CTHRC1, MMP2 and FUNDC1 mRNA expression in ovarian cancer tissue were significantly higher than those in adjacent tissue whereas NDRG1, E-cadherin and Wnt5a mRNA expression were significantly lower than those in adjacent tissue;YKL40, COX2, Twist, IFITM1, CatL, CTHRC1, MMP2 and FUNDC1 mRNA expression in ovarian cancer tissue with high LSD1 expression were significantly higher than those in ovarian tissue with low LSD1 expression whereas E-cadherin and Wnt5a gene mRNA expression were significantly lower than those in ovarian tissue with low LSD1 expression. Conclusion: The high LSD1 expression and low NDRG1 expression in ovarian cancer tissue can promote the migration and invasion of cancer cells.

Correlation of LSD1 and PARP1 with cell proliferation and epithelial-mesenchymal transition in ovarian cancer tissue from ultrasound-guided puncture

Objective: To study the correlation of LSD1 and PARP1 with cell proliferation and epithelial-mesenchymal transition in ovarian cancer tissue from ultrasound-guided puncture. Methods:The ovarian cancer and ovarian benign lesion tissue from ultrasound-guided puncture in Pangang Group General Hospital in Panzhihua between May 2014 and March 2017 were collected to detect the mRNA expression of LSD1 and PARP1 as well as the protein levels of cell proliferation molecules and epithelial-mesenchymal transition molecules in them. Results: LSD1 and PARP1 mRNA expression in ovarian cancer tissue were significantly higher than those in benign ovarian tissue;P21, P27 and E-cadherin protein levels in ovarian cancer tissue were significantly lower than those in benign ovarian tissue while CyclinD1, E2F, Twist1, Snail, Slug and N-cadherin protein levels were significantly higher those in benign ovarian tissue;P21 and P27 protein levels in the ovarian cancer tissue with high LSD1 expression were significantly lower than those in the ovarian cancer tissue with low LSD1 expression while CyclinD1 and E2F protein levels were significantly higher than those in the ovarian cancer tissue with low LSD1 expression;Twist1, Snail, Slug and N-cadherin protein levels in the ovarian cancer tissue with high PARP1 expression were significantly higher than those in the ovarian cancer tissue with low PARP1 expression while E-cadherin protein level was significantly lower than that in the ovarian cancer tissue with low PARP1 expression. Conclusion: The LSD1 and PARP1 highly expressed in ovarian cancer tissue can promote the proliferation and epithelial-mesenchymal transition of cancer cells.

Targeting histone lysine-specific demethylase KDM1A/LSD1 to control epithelial-mesenchymal transition program in breast cancers

Epithelial-mesenchymal transition (EMT) is a plastic and reversible process, essential for development and tissue homeostasis. Under pathological conditions, EMT causes induction of tumor growth, angiogenesis and metastasis. According to its reversible nature, the EMT program is associated with vast epigenetic changes. Targeting the epigenetic network that controls the EMT pathway in disease progression is a novel promising strategy to fight cancer metastasis. The impact of alterations in histone methylation in cancer has led to the identification of histone methyltransferases and demethylases as promising novel targets for therapy. Specifically, the lysine specific demethylase 1 (LSD1, also known as KDM1A) plays a pivotal role in the regulation of EMT. Here we present an overview of the causative role of LSD1 in the EMT process, summarizing recent findings on its emerging functions in cell migration and invasion in breast cancer.

Modulation of cell death pathways in cancer stem cells: Targeting histone demethylases

Cancer stem cells (CSCs) are tumor initiating cells within the tumor mass;that play a critical role in cancer pathogenesis. CSCs regulate cancer cell survival, metastatic potential, resistance to conventional radio-chemotherapy, disease relapse and poor prognosis. Recent studies have established that the drug resistant cancers and cancer cell lines possess high stem cell like traits compared to their drug sensitive counterparts. Histone demethylases are recently been linked to drug induced reversible tolerant state in cancers. Lysine histone demethylases are enzymes those demethylate lysines in histones and can act as transcriptional repressors or activators. Apart from histones other cellular proteins like E2F1, Rb, STAT3 and p53 are also regulated by methylation and demethylation cycles. In cancer cells these enzymes regulate cell survival, migration, invasion, and proliferation. This review summarizes the current progress of research on the role of histone demethylases in supporting drug tolerant cancer stem cell state and their potential as a drug target.

Development of the triazole-fused pyrimidine derivatives as highly potent and reversible inhibitors of histone lysine specific demethylase1(LSD1/KDM1A)

Histone lysine specific demethylase 1(LSD1) has been recognized as an important modulator in post-translational process in epigenetics. Dysregulation of LSD1 has been implicated in the development of various cancers. Herein, we report the discovery of the hit compound 8 a(IC50=3.93 μmol/L) and further medicinal chemistry efforts, leading to the generation of compound 15 u(IC50=49 nmol/L, and Ki= 16 nmol/L), which inhibited LSD1 reversibly and competitively with H3 K4 me2, and was selective to LSD1 over MAO-A/B. Docking studies were performed to rationalize the potency ofcompound 15 u. Compound 15 u also showed strong antiproliferative activity against four leukemia cell lines(OCL-AML3, K562, THP-1 and U937) as well as the lymphoma cell line Raji with the IC50 values of 1.79, 1.30, 0.45, 1.22 and 1.40 μmol/L, respectively. In THP-1 cell line, 15 u significantly inhibited colony formation and caused remarkable morphological changes. Compound 15 u induced expression of CD86 and CD11 b in THP-1 cells, confirming its cellular activity and ability of inducing differentiation.The findings further indicate that targeting LSD1 is a promising strategy for AML treatment, the triazolefused pyrimidine derivatives are new scaffolds for the development of LSD1/KDM1 A inhibitors.

Promising natural lysine specific demethylase 1 inhibitors for cancer treatment:advances and outlooks

Lysine specific demethylase 1(LSD1),a transcriptional corepressor or coactivator that serves as a demethylase of histone 3 lysine 4 and 9,has become a potential therapeutic target for cancer therapy.LSD1 mediates many cellular signaling pathways and regulates cancer cell proliferation,invasion,migration,and differentiation.Recent research has focused on the exploration of its pharmacological inhibitors.Natural products are a major source of compounds with abundant scaffold diversity and structural complexity,which have made a major contribution to drug discovery,particularly anticancer agents.In this review,we briefly highlight recent advances in natural LSD1 inhibitors over the past decade.We present a comprehensive review on their discovery and identification process,natural plant sources,chemical structures,anticancer effects,and structure-activity relationships,and finally provide our perspective on the development of novel natural LSD1 inhibitors for cancer therapy.

Natural products as LSD1 inhibitors for cancer therapy

Natural products generally fall into the biologically relevant chemical space and always possess novel biological activities, thus making them a rich source of lead compounds for new drug discovery. With the recent technological advances, natural product-based drug discovery is now reaching a new era. Natural products have also shown promise in epigenetic drug discovery, some of them have advanced into clinical trials or are presently being used in clinic. The histone lysine specific demethylase1(LSD1), an important class of histone demethylases, has fundamental roles in the development of various pathological conditions. Targeting LSD1 has been recognized as a promising therapeutic option for cancer treatment. Notably, some natural products with different chemotypes including protoberberine alkaloids, flavones, polyphenols, and cyclic peptides have shown effectiveness against LSD1. These natural products provide novel scaffolds for developing new LSD1 inhibitors. In this review, we mainly discuss the identification of natural LSD1 inhibitors, analysis of the co-crystal structures of LSD1/natural product complex, antitumor activity and their modes of action. We also briefly discuss the challenges faced in this field. We believe this review will provide a landscape of natural LSD1 inhibitors.

赖氨酸特异性组蛋白去甲基化酶1在肿瘤中的研究进展

赖氨酸特异性组蛋白去甲基化酶1(LSD1)是第一个被发现的组蛋白去甲基化酶,调控基因的转录和p53的活性,在多种肿瘤组织中高表达,与癌症的发生发展高度相关。全文对近年来LSD1在肿瘤中的最新研究进展进行综述。

组蛋白去甲基化酶的研究进展

第一个组蛋白去甲基化酶(LSD1)的发现证实组蛋白的甲基化修饰并非永久性组蛋白标记,而是一个动态可调节过程。该酶通过催化去除组蛋白N末端赖氨酸上的甲基,参与染色质功能的调节,并与某些疾病,特别是和癌症的发生发展密切相关。本文主要就组蛋白去甲基化酶的催化机制及生物学功能进行文献调研与追踪,并加以归纳整理,旨在更深入的认识和探究组蛋白去甲基化酶提供理论参考。