Micropeptides are endogenous peptides translated from alternative open reading frames (alt-ORFs) within coding or non-coding genes. Emerging evidence suggests that some micropeptides play critical roles in both physiological and pathological processes. Multiple myeloma (MM), a haematological malignancy, remains incurable due to frequent relapses and a limited understanding of its underlying mechanisms. In this study, we sought to investigate the function and molecular mechanism of a novel micropeptide in MM pathogenesis. We identified a novel micropeptide, altH19, encoded by the lncRNA H19, which is highly expressed in patients of MM. Functional assays revealed that altH19 promotes myeloma cell proliferation and colony formation significantly. Furthermore, altH19 induces multipolar mitosis by upregulating the expression of Aurora B, Centrin 2 and phosphorylated histone H3. Flow cytometry analyses confirmed that overexpression of altH19 enhances DNA replication and accelerates the transition from early to mid-late stages of the DNA replication process. Conversely, knockout of altH19 reverses these effects. Mechanistically, altH19 directly interacts with phosphorylated CDK2 at threonine 160, thereby enhancing CDK2 T160 phosphorylation and activating the downstream E2F1 target RB phosphorylation. Notably, altH19 was able to restore phosphorylation levels of CDK2 and RB that were otherwise suppressed by the CDK2-selective inhibitor Seliciclib. In summary, we identify altH19 as a novel lncRNA-derived micropeptide with a pivotal role in myeloma progression, highlighting the therapeutic potential of targeting the altH19-CDK2-RB axis in MM treatment.
| [1] |
X. Cao, A. Khitun, Z. Na, et al., “Comparative Proteomic Profiling of Unannotated Microproteins and Alternative Proteins in Human Cell Lines,” Journal of Proteome Research 19, no. 8 (2020): 3418–3426.
|
| [2] |
R. Benezra, R. L. Davis, D. Lockshon, D. L. Turner, and H. Weintraub, “The Protein Id: A Negative Regulator of Helix-Loop-Helix DNA Binding Proteins,” Cell 61, no. 1 (1990): 49–59.
|
| [3] |
S. Azam, F. Yang, and X. Wu, “Finding Functional Microproteins,” Trends in Genetics 41, no. 2 (2025): 107–118.
|
| [4] |
N. G. D'Lima, J. Ma, L. Winkler, et al., “A Human Microprotein That Interacts With the mRNA Decapping Complex,” Nature Chemical Biology 13, no. 2 (2017): 174–180.
|
| [5] |
R. Yu, Y. Hu, S. Zhang, et al., “LncRNA CTBP1-DT-Encoded Microprotein DDUP Sustains DNA Damage Response Signalling to Trigger Dual DNA Repair Mechanisms,” Nucleic Acids Research 50, no. 14 (2022): 8060–8079.
|
| [6] |
A. Chugunova, E. Loseva, P. Mazin, et al., “LINC00116 Codes for a Mitochondrial Peptide Linking Respiration and Lipid Metabolism,” Proceedings of the National Academy of Sciences of the United States of America 116, no. 11 (2019): 4940–4945.
|
| [7] |
Q. Zhang, A. A. Vashisht, J. O'Rourke, et al., “The Microprotein Minion Controls Cell Fusion and Muscle Formation,” Nature Communications 8 (2017): 15664.
|
| [8] |
L. Ren, X. Qing, J. Wei, et al., “The DDUP Protein Encoded by the DNA Damage-Induced CTBP1-DT lncRNA Confers Cisplatin Resistance in Ovarian Cancer,” Cell Death & Disease 14, no. 8 (2023): 568.
|
| [9] |
M. Polycarpou-Schwarz, M. Gross, P. Mestdagh, et al., “The Cancer-Associated Microprotein CASIMO1 Controls Cell Proliferation and Interacts With Squalene Epoxidase Modulating Lipid Droplet Formation,” Oncogene 37, no. 34 (2018): 4750–4768.
|
| [10] |
W. Xu, B. Deng, P. Lin, et al., “Ribosome Profiling Analysis Identified a KRAS-Interacting Microprotein That Represses Oncogenic Signaling in Hepatocellular Carcinoma Cells,” Science China Life Sciences 63, no. 4 (2020): 529–542.
|
| [11] |
M. Zhang, K. Zhao, X. Xu, et al., “A Peptide Encoded by Circular Form of LINC-PINT Suppresses Oncogenic Transcriptional Elongation in Glioblastoma,” Nature Communications 9, no. 1 (2018): 4475.
|
| [12] |
Q. Ge, D. Jia, D. Cen, et al., “Micropeptide ASAP Encoded by LINC00467 Promotes Colorectal Cancer Progression by Directly Modulating ATP Synthase Activity,” Journal of Clinical Investigation 131, no. 22 (2021): e152911.
|
| [13] |
Y. Godet, A. Moreau-Aubry, D. Mompelat, et al., “An Additional ORF on Meloe cDNA Encodes a New Melanoma Antigen, MELOE-2, Recognized by Melanoma-Specific T Cells in the HLA-A2 Context,” Cancer Immunology, Immunotherapy: CII 59, no. 3 (2010): 431–439.
|
| [14] |
L. Sun, W. Wang, C. Han, et al., “The Oncomicropeptide APPLE Promotes Hematopoietic Malignancy by Enhancing Translation Initiation,” Molecular Cell 81, no. 21 (2021): 4493–4508.
|
| [15] |
R. Yao, Y. Zeng, Y. Zhang, et al., “Identification of a New Micropeptide altKLF4 Derived From KLF4 That Influences Myeloma Chemotherapeutic Sensitivity,” Cellular Signalling 131 (2025): 111767.
|
| [16] |
S. K. Kumar, V. Rajkumar, R. A. Kyle, et al., “Multiple Myeloma,” Nature Reviews Disease Primers 3 (2017): 17046.
|
| [17] |
U. H. Weidle, F. Birzele, G. Kollmorgen, and R. Ruger, “Molecular Mechanisms of Bone Metastasis,” Cancer Genomics & Proteomics 13, no. 1 (2016): 1–12.
|
| [18] |
S. V. Glavey, S. Manier, A. Sacco, et al., “Epigenetics in Multiple Myeloma,” Cancer Treatment and Research 169 (2016): 35–49.
|
| [19] |
N. van de Donk, C. Pawlyn, and K. L. Yong, “Multiple Myeloma,” Lancet 397, no. 10272 (2021): 410–427.
|
| [20] |
J. Abraham, N. N. Salama, and A. K. Azab, “The Role of P-Glycoprotein in Drug Resistance in Multiple Myeloma,” Leukemia & Lymphoma 56, no. 1 (2015): 26–33.
|
| [21] |
G. J. Morgan, B. A. Walker, and F. E. Davies, “The Genetic Architecture of Multiple Myeloma,” Nature Reviews Cancer 12, no. 5 (2012): 335–348.
|
| [22] |
M. Malumbres and M. Barbacid, “Cell Cycle, CDKs and Cancer: A Changing Paradigm,” Nature Reviews Cancer 9, no. 3 (2009): 153–166.
|
| [23] |
R. Fagundes and L. K. Teixeira, “Cyclin E/CDK2: DNA Replication, Replication Stress and Genomic Instability,” Frontiers in Cell and Developmental Biology 9 (2021): 774845.
|
| [24] |
S. Ghafouri-Fard, M. Esmaeili, and M. Taheri, “H19 lncRNA: Roles in Tumorigenesis,” Biomedicine & Pharmacotherapy 123 (2020): 109774.
|
| [25] |
J. F. Zheng, N. H. Guo, F. M. Zi, and J. Cheng, “Long Noncoding RNA H19 Promotes Tumorigenesis of Multiple Myeloma by Activating BRD4 Signaling by Targeting MicroRNA 152-3p,” Molecular and Cellular Biology 40, no. 3 (2020): e00382-19.
|
| [26] |
F. Zhan, Y. Huang, S. Colla, et al., “The Molecular Classification of Multiple Myeloma,” Blood 108, no. 6 (2006): 2020–2028.
|
| [27] |
G. Maryu and Q. Yang, “Nuclear-Cytoplasmic Compartmentalization of Cyclin B1-Cdk1 Promotes Robust Timing of Mitotic Events,” Cell Reports 41, no. 13 (2022): 111870.
|
| [28] |
T. Abbas, S. Jha, N. E. Sherman, and A. Dutta, “Autocatalytic Phosphorylation of CDK2 at the Activating Thr160,” Cell Cycle 6, no. 7 (2007): 843–852.
|
| [29] |
S. W. Choi, H. W. Kim, and J. W. Nam, “The Small Peptide World in Long Noncoding RNAs,” Briefings in Bioinformatics 20, no. 5 (2019): 1853–1864.
|
| [30] |
Y. Xia, T. Pei, J. Zhao, et al., “Long Noncoding RNA H19: Functions and Mechanisms in Regulating Programmed Cell Death in Cancer,” Cell Death Discovery 10, no. 1 (2024): 76.
|
| [31] |
J. Wang, S. Xie, J. Yang, et al., “The Long Noncoding RNA H19 Promotes Tamoxifen Resistance in Breast Cancer via Autophagy,” Journal of Hematology & Oncology 12, no. 1 (2019): 81.
|
| [32] |
P. Wan, W. Su, Y. Zhang, et al., “LncRNA H19 Initiates Microglial Pyroptosis and Neuronal Death in Retinal Ischemia/Reperfusion Injury,” Cell Death and Differentiation 27, no. 1 (2020): 176–191.
|
| [33] |
D. Coverley, H. Laman, and R. A. Laskey, “Distinct Roles for Cyclins E and A During DNA Replication Complex Assembly and Activation,” Nature Cell Biology 4, no. 7 (2002): 523–528.
|
| [34] |
T. M. Guadagno and J. W. Newport, “Cdk2 Kinase Is Required for Entry Into Mitosis as a Positive Regulator of Cdc2-Cyclin B Kinase Activity,” Cell 84, no. 1 (1996): 73–82.
|
| [35] |
D. Gisselsson, T. Jonson, C. Yu, et al., “Centrosomal Abnormalities, Multipolar Mitoses, and Chromosomal Instability in Head and Neck Tumours With Dysfunctional Telomeres,” British Journal of Cancer 87, no. 2 (2002): 202–207.
|
| [36] |
N. J. Ganem, S. A. Godinho, and D. Pellman, “A Mechanism Linking Extra Centrosomes to Chromosomal Instability,” Nature 460, no. 7252 (2009): 278–282.
|
| [37] |
S. Manier, K. Z. Salem, J. Park, D. A. Landau, G. Getz, and I. M. Ghobrial, “Genomic Complexity of Multiple Myeloma and Its Clinical Implications,” Nature Reviews Clinical Oncology 14, no. 2 (2017): 100–113.
|
RIGHTS & PERMISSIONS
2025 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.
PDF
(5374KB)
