A novel protein encoded by circHNRNPU promotes multiple myeloma progression by regulating the bone marrow microenvironment and alternative splicing.
A novel protein encoded by circHNRNPU promotes multiple myeloma progression by regulating the bone marrow microenvironment and alternative splicing.
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circHNRNPU编码的新型蛋白质通过调节骨髓微环境和选择性剪接促进多发性骨髓瘤进展
DOI：
10.1186/s13046-022-02276-7
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发表时间：
2022-03-08
期刊：
影响因子：
--
通讯作者：
Gu C
中科院分区：
文献类型：
--
作者：
Tang X;Deng Z;Ding P;Qiang W;Lu Y;Gao S;Hu Y;Yang Y;Du J;Gu C
多发性骨髓瘤（MM）是骨髓（BM）中不可治愈的浆细胞恶性肿瘤，而免疫球蛋白D型MM（IgD MM）是所有MM病例中非常罕见但最严重的亚型。因此，对IgD MM进行系统研究，有助于揭示IgD和其他类型MM的复发和难治特征，并有助于MM有效治疗策略的制定。采用Agilent SBC-ceRNA微阵列芯片分别检测3例正常浆细胞（NPC）、5例IgD MM和5例IgG MM。桑格测序、RNase R酶切和qPCR检测circHNRNPU的存在和表达。进行BaseScope™ RNA ISH测定以测试石蜡包埋的MM组织中的circHNRNPU水平。经LC-MS/MS鉴定，编码的蛋白为circHNRNPU_603aa。采用MTT法、集落形成实验、流式细胞术和MM小鼠移植瘤模型检测circHNRNPU_603aa对细胞增殖和细胞周期的影响。通过RIP-seq、RIP-PCR和WB分析泛素化以探索circHNRNPU_603 aa在MM中的潜在机制。通过超离心从MM细胞的培养上清液中分离外泌体，并通过透射电子显微镜和WB确认外泌体标记物阿利克斯和CD 9来表征。相对于IgG和NPC样品，CircHNRNPU是IgD MM中最丰富和差异表达的circRNA之一。在4个独立的MM患者队列中，circHNRNPU增加与不良结局相关。有趣的是，MM细胞分泌circHNRNPU，它编码一种名为circHNRNPU_603aa的蛋白质。过表达的circHNRNPU_603aa在体外和体内促进MM细胞增殖，相反，通过siRNA敲低circHNRNPU_603aa消除了这些作用。由于circHNRNPU_603aa包含RNA结合的RGG盒区域，它调节SKP 2外显子跳跃，从而竞争性抑制c-Myc泛素，从而稳定MM中的c-Myc。MM细胞通过外泌体分泌circHNRNPU，干扰BM微环境中的各种细胞。我们的研究结果表明，circHNRNPU_603aa是MM细胞和BM龛中有前途的诊断和治疗标志物。在线版本包含补充材料，可通过10.1186/s13046-022-02276-7获得。
Multiple myeloma (MM) is an incurable plasma cell malignancy in the bone marrow (BM), while immunoglobulin D type of MM (IgD MM) is a very rare but most severe subtype in all MM cases. Therefore, systemic study on IgD MM is purposeful to disclose the recurrent and refractory features in both IgD and other types of MM, and beneficial to the development of potent therapeutic strategy on MM. Agilent SBC-ceRNA microarray chips were employed to examine 3 normal plasma cell samples (NPCs), 5 lgD MM samples and 5 lgG MM samples, respectively. Sanger sequencing, RNase R digestion and qPCR assays were used to detect the existence and expression of circHNRNPU. BaseScope™ RNA ISH assay was performed to test circHNRNPU levels in paraffin-embedded MM tissues. The protein encoded by circHNRNPU was identified by LC-MS/MS, which was named as circHNRNPU_603aa. The function of circHNRNPU_603aa on cellular proliferation and cell cycle was assessed by MTT test, colony formation assay, flow cytometry and MM xenograft mouse model in vivo. RIP-seq, RIP-PCR and WB analysis for ubiquitination were performed to explore the potential mechanism of circHNRNPU_603aa in MM. Exosomes were isolated from the culture supernatant of MM cells by ultracentrifugation and characterized by Transmission Electron Microscope and WB confirmation of exosomes markers Alix and CD9. CircHNRNPU was one of the top most abundant and differentially expressed circRNA in IgD MM relative to lgG and NPCs samples. Increased circHNRNPU was associated with poor outcomes in four independent MM patient cohorts. Intriguingly, MM cells secreted circHNRNPU, which encoded a protein named as circHNRNPU_603aa. Overexpressed circHNRNPU_603aa promoted MM cell proliferation in vitro and in vivo, in contrast knockdown of circHNRNPU_603aa by siRNA abrogated these effects. Due to circHNRNPU_603aa including RNA-binding RGG-box region, it regulated SKP2 exon skipping, thereby competitively inhibited c-Myc ubiquitin so as to stabilize c-Myc in MM. MM cells secreted circHNRNPU through exosomes to interfere with various cells in the BM microenvironment. Our findings demonstrate that circHNRNPU_603aa is a promising diagnostic and therapeutic marker in both MM cells and BM niche. The online version contains supplementary material available at 10.1186/s13046-022-02276-7.
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影响因子：
64.5
作者：
Cowan CS;Renner M;De Gennaro M;Gross-Scherf B;Goldblum D;Hou Y;Munz M;Rodrigues TM;Krol J;Szikra T;Cuttat R;Waldt A;Papasaikas P;Diggelmann R;Patino-Alvarez CP;Galliker P;Spirig SE;Pavlinic D;Gerber-Hollbach N;Schuierer S;Srdanovic A;Balogh M;Panero R;Kusnyerik A;Szabo A;Stadler MB;Orgül S;Picelli S;Hasler PW;Hierlemann A;Scholl HPN;Roma G;Nigsch F;Roska B
通讯作者：
Roska B
影响因子：
4.6
作者：
Abe N;Matsumoto K;Nishihara M;Nakano Y;Shibata A;Maruyama H;Shuto S;Matsuda A;Yoshida M;Ito Y;Abe H
通讯作者：
Abe H
影响因子：
14.5
作者：
Cai, Zhen;Moten, Asad;Peng, Danni;Hsu, Che-Chia;Pan, Bo-Syong;Manne, Rajeshkumar;Li, Hong-yu;Lin, Hui-Kuan
通讯作者：
Lin, Hui-Kuan
影响因子：
45.3
作者：
BLADE, J;LUST, JA;KYLE, RA
通讯作者：
KYLE, RA
影响因子：
20.3
作者：
Keats, Jonathan J.;Chesi, Marta;Bergsagel, P. Leif
通讯作者：
Bergsagel, P. Leif