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GFI-1 and Osteoblast Suppression in Multiple Myeloma

多发性骨髓瘤中的 GFI-1 和成骨细胞抑制

基本信息

批准号：
8681152
负责人：
DEBORAH Lynn GALSON
金额：
$ 27.21万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-26 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8681152
关键词：
Ascorbic Acid BMP2 gene Binding Binding Sites Bone Diseases Bone Pain Cell Differentiation process Cells Cessation of life Clinical Detection Differentiation Antigens Disease remission Embryo Environment Enzymes Epigenetic Process Fracture Ganciclovir Gene Expression Genes Growth Healed Histones In Vitro Interleukin-6 Interleukin-7 Lesion Lytic Lytic Metastatic Lesion Maintenance Malignant Neoplasms Mediating Mesenchymal Multiple Myeloma Mus Osteoblasts Osteoclasts Osteogenesis Pathological fracture Patients Plasmids Play Production Recruitment Activity Reporting Repression Risk Role Skeleton Small Interfering RNA Stromal Cells TNF gene TNFSF11 gene Testing Thymidine Kinase Transcription Repressor/Corepressor Transfection Tumor Necrosis Factor Suppression Tumor Necrosis Factor-alpha Up-Regulation base bone cell growth chromatin remodeling deletion analysis economic impact healing in vivo in vivo Model killings lipid biosynthesis mouse model osteoblast differentiation prevent promoter transcription factor

项目摘要

DESCRIPTION (provided by applicant): Myeloma (MM) is the most frequent cancer to involve the skeleton. Up to 90% of patients develop bone lesions that can result in severe bone pain and frequent pathologic fractures. Unfortunately, these bone lesions rarely heal even when patients are in long-term remission because of the permanent MM-induced suppression of osteoblast precursor (OBP) differentiation into functional bone-forming osteoblasts. MM induces an intrinsic and persistent change in the OBP differentiation potential, the basis of which is unknown. Preliminary studies with a murine in vivo model of MM-induced OB suppression using 5TGM1 MM cells (which we modified to express GFP for detection and thymidine kinase, which allows them to be selectively killed by ganciclovir) demonstrate a persistent inhibition of OB differentiation even in the absence of MM cells. The OBP from these mice maintained low levels of the critical OB transcription factor, Runx2 even when induced to differentiate and had elevated expression of the transcriptional repressor Gfi-1, which can mediate chromatin remodeling. Importantly, Gfi-1 levels in OBP from 7/7 MM patients were elevated compared to 3 normals. 5TGM1 MM cells inhibited OB differentiation in vitro by producing TNF-1 and IL-7, which increased Gfi-1 in a mouse OBP line (MC4). Further, mouse Runx2 promoter analysis identified a 1003 bp region (-992/+111) that is responsible for suppression of Runx2 expression by MM cells and contains 29 putative Gfi-1 binding sites. This region is also repressed by TNF-1 and by co-transfection with a Gfi-1 expression plasmid. Importantly, knockdown of Gfi-1 expression, using a specific siRNA, significantly restored expression of Runx2 as well as the expression of several OB markers in both MC4 cells pretreated with MM cells and in MSC from 2 MM patients. These results suggest the hypothesis that MM cells secrete soluble factors (TNF-1 and IL-7) that increase Gfi-1 expression in OBPs. Gfi-1 then suppresses Runx2 production and thereby inhibits osteoblastogenesis. Further, since Gfi-1 can recruit to genes histone-modifying enzymes that create epigenetic changes, this results in long-term suppression of Runx2 that is maintained in the absence of MM cells. However, the role of Gfi-1 plays in OBP differentiation and in MM in particular is currently unknown and is the focus of this proposal. The following specific aims will be pursued to test this hypothesis: (1) Determine if Gfi-1 up-regulation in MSC is necessary for MM or TNF-1/IL-7 suppression of OB differentiation. (2) Determine if elevated Gfi-1 is sufficient for OB suppression and/or for increased IL-6 and RANKL production by MSC and does it act by direct binding to the Runx2 gene. (3) Determine if MM cells induce epigenetic changes in the Runx2 gene in MSC via Gfi-1, and if they are responsible for long-term OB suppression by assessing if altering the epigenetic status of the Runx2 gene relieves the differentiation block. (4) Determine if Gfi-1 deficiency in MSC in vivo prevents MM-induced suppression of OB differentiation.

描述(申请人提供)：骨髓瘤(MM)是最常见的累及骨骼的癌症。高达90%的患者出现骨损伤，可能导致严重的骨痛和频繁的病理性骨折。不幸的是，这些骨损伤即使在患者长期缓解的情况下也很少愈合，因为MM诱导的成骨细胞前体(OBP)向功能性成骨细胞分化的永久性抑制。MM引起OBP分化潜能的内在和持久的改变，其基础尚不清楚。使用5TGM1 MM细胞(我们将其修饰为表达用于检测的GFP和胸苷激酶，使其能够选择性地被更昔洛韦杀死)抑制MM诱导的OB分化的小鼠体内模型的初步研究表明，即使没有MM细胞，OB分化也会持续受到抑制。这些小鼠的OBP即使在被诱导分化时仍保持着关键的OB转录因子Runx2的低水平，并上调了转录抑制因子GFI-1的表达，GFI-1可以介导染色质的重塑。重要的是，7/7的MM患者OBP中的GFI-1水平高于3名正常人。5TGM1 MM细胞体外通过产生肿瘤坏死因子-1和白介素7抑制成骨细胞的分化，使小鼠成骨细胞系(MC4)中的GFI-1增加。此外，小鼠Runx2启动子分析确定了一个1003bp的区域(-992/+111)，该区域负责抑制MM细胞Runx2的表达，并包含29个推测的GFI-1结合位点。这一区域也被肿瘤坏死因子-1和与GFI-1表达载体共转染而抑制。重要的是，利用特定的siRNA抑制GFI-1的表达，显著恢复了经MM细胞处理的MC4细胞和2例MM患者的MSC中Runx2的表达以及几个OB标记的表达。这些结果提示MM细胞分泌的可溶性因子(TNF-1和IL-7)可增加OBP中GFI-1的表达。然后GFI-1抑制Runx2的产生，从而抑制成骨细胞的形成。此外，由于GFI-1可以招募产生表观遗传变化的组蛋白修饰酶基因，这导致了对Runx2的长期抑制，这种抑制在没有MM细胞的情况下保持不变。然而，GFI-1在OBP分化中的作用，特别是在MM中的作用目前尚不清楚，这是本提案的重点。为了验证这一假说，将寻求以下具体目标：(1)确定MSC中GFI-1的上调是否是MM或TNF-1/IL-7抑制OB分化所必需的。(2)确定升高的GFI-1是否足以抑制OB和/或增加MSC产生的IL-6和RANKL，以及它是否通过直接与Runx2基因结合起作用。(3)确定MM细胞是否通过GFI-1诱导MSC中Runx2基因的表观遗传学改变，并通过评估改变Runx2基因的表观遗传状态是否解除分化障碍来确定它们是否对长期的OB抑制负责。(4)确定体内骨髓间充质干细胞中GFI-1缺乏是否能阻止MM诱导的OB分化抑制。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Resveratrol triggers the pro-apoptotic endoplasmic reticulum stress response and represses pro-survival XBP1 signaling in human multiple myeloma cells.

白藜芦醇触发促凋亡的内质网应激反应，并抑制人类多发性骨髓瘤细胞中的促生物性XBP1信号传导。

DOI：
10.1016/j.exphem.2011.06.007
发表时间：
2011-10
期刊：
EXPERIMENTAL HEMATOLOGY
影响因子：
2.6
作者：
Wang, Feng-Ming;Galson, Deborah L.;Roodman, G. David;Ouyang, Hongjiao
通讯作者：
Ouyang, Hongjiao

Mechanisms of multiple myeloma bone disease.

DOI：
10.1038/bonekey.2012.135
发表时间：
2012-01-01
期刊：
BoneKEy reports
影响因子：
0
作者：
Galson, Deborah L;Silbermann, Rebecca;Roodman, G David
通讯作者：
Roodman, G David

GFI1-Dependent Repression of SGPP1 Increases Multiple Myeloma Cell Survival.

DOI：
10.3390/cancers14030772
发表时间：
2022-02-02
期刊：
Cancers
影响因子：
5.2
作者：
Petrusca DN;Mulcrone PL;Macar DA;Bishop RT;Berdyshev E;Suvannasankha A;Anderson JL;Sun Q;Auron PE;Galson DL;Roodman GD
通讯作者：
Roodman GD

XRK3F2 Inhibition of p62-ZZ Domain Signaling Rescues Myeloma-Induced GFI1-Driven Epigenetic Repression of the Runx2 Gene in Pre-osteoblasts to Overcome Differentiation Suppression.

DOI：
10.3389/fendo.2018.00344
发表时间：
2018
期刊：
Frontiers in endocrinology
影响因子：
5.2
作者：
Adamik J;Silbermann R;Marino S;Sun Q;Anderson JL;Zhou D;Xie XQ;Roodman GD;Galson DL
通讯作者：
Galson DL

The Role of Semaphorin 4D in Bone Remodeling and Cancer Metastasis.