Abstract
Background
Mesenchymal stem cells (MSCs) play an important role in regulating all stages of the immune response, angiogenesis, and transformation of matrix components in the tumor microenvironment. The aim of this study was to identify the prognostic value of MSC-related signatures in patients with gastric cancer (GC).
Methods
MSC marker genes were identified by analyzing single-cell RNA sequencing (scRNA-seq) data for GC from the Gene Expression Omnibus (GEO) database. Using bulk sequencing data from the Cancer Genome Atlas-Stomach adenocarcinoma (TCGA-STAD), as a training cohort, and data from GEO, as a validation cohort, we developed a risk model consisting of MSC prognostic signature genes, and classified GC patients into high- and low-MSC risk subgroups. Multifactorial Cox regression was used to evaluate whether MSC prognostic signature was an independent prognostic factor. An MSC nomogram was constructed combining clinical information and risk grouping. Subsequently, we evaluated the effect of MSC prognostic signature on immune cell infiltration, antitumor drugs and immune checkpoints and verified the expression of MSC prognostic signature by in vitro cellular assays.
Results
In this study, 174 MSC marker genes were identified by analyzing scRNA-seq data. We identified seven genes (POSTN, PLOD2, ITGAV, MMP11, SDC2, MARCKS, ANXA5) to construct MSC prognostic signature. MSC prognostic signature was an independent risk factor in the TCGA and GEO cohorts. GC patients in the high-MSC risk group had worse prognoses. In addition, the MSC nomogram has a high clinical application value. Notably, the MSC signature can induce the development of a poor immune microenvironment. GC patients in the high MSC-risk group were more sensitive to anticancer drugs and tended to have higher levels of immune checkpoint markers. In qRT-PCR assays, the MSC signature was more highly expressed in GC cell lines.
Conclusions
The MSC marker gene-based risk signature developed in this study can not only be used to predict the prognosis of GC patients, but also has the potential to reflect the efficacy of antitumor therapies
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The scRNA-seq(GSE163558) and bulk sequencing(GSE15459) data from this study are available in the GEO (https://www.ncbi.nlm.nih.gov/geo/) and TGCA (https://portal.gdc.cancer.gov/) databases,other data were obtained from corresponding authors upon reasonable request.
References
Ahn J, Lee H, Seo K et al (2013) Anti-tumor effect of adipose tissue derived-mesenchymal stem cells expressing interferon-β and treatment with cisplatin in a xenograft mouse model for canine melanoma. PLoS ONE 8(9):e74897. https://doi.org/10.1371/journal.pone.0074897
Anderson LA, Tavilla A, Brenner H et al (2007) Survival for oesophageal, stomach and small intestine cancers in Europe 1999–2007: results from EUROCARE-5. Eur J Cancer 51(15):2144–2157. https://doi.org/10.1016/j.ejca.2015.07.026
Aravindhan S, Ejam SS, Lafta MH et al (2021) Mesenchymal stem cells and cancer therapy: insights into targeting the tumour vasculature. Cancer Cell Int 21(1):158. https://doi.org/10.1186/s12935-021-01836-9
Arrington CB, Yost HJ (2009) Extra-embryonic syndecan 2 regulates organ primordia migration and fibrillogenesis throughout the zebrafish embryo. Development 136(18):3143–3152. https://doi.org/10.1242/dev.031492
Balachandran VP, Gonen M, Smith JJ, DeMatteo RP (2015) Nomograms in oncology: more than meets the eye. Lancet Oncol 16(4):e173-180. https://doi.org/10.1016/S1470-2045(14)71116-7
Bang YJ, Ruiz EY, Van Cutsem E et al (2018) Phase III, randomised trial of avelumab versus physician’s choice of chemotherapy as third-line treatment of patients with advanced gastric or gastro-oesophageal junction cancer: primary analysis of JAVELIN Gastric 300. Ann Oncol 29(10):2052–2060. https://doi.org/10.1093/annonc/mdy264
Bhattarai P, Hameed S, Dai Z (2018) Recent advances in anti-angiogenic nanomedicines for cancer therapy. Nanoscale 10(12):5393–5423. https://doi.org/10.1039/c7nr09612g
Charbord P (2010) Bone marrow mesenchymal stem cells: historical overview and concepts. Hum Gene Ther 21(9):1045–1056. https://doi.org/10.1089/hum.2010.115
Chen CH, Statt S, Chiu CL et al (2014a) Targeting myristoylated alanine-rich C kinase substrate phosphorylation site domain in lung cancer, mechanisms and therapeutic implications. Am J Respir Crit Care Med 190(10):1127–1138. https://doi.org/10.1164/rccm.201408-1505OC
Chen CH, Thai P, Yoneda K et al (2014b) A peptide that inhibits function of myristoylated alanine-rich C kinase substrate (MARCKS) reduces lung cancer metastasis. Oncogene 33(28):3696–3706. https://doi.org/10.1038/onc.2013.336
Chen CH, Fong LWR, Yu E et al (2016) Upregulation of MARCKS in kidney cancer and its potential as a therapeutic target. Oncogene 36(25):3588–3598. https://doi.org/10.1038/onc.2016.510
Choi S, Kim Y, Park H et al (2009) Syndecan-2 overexpression regulates adhesion and migration through cooperation with integrin alpha2. Biochem Biophys Res Commun 384(2):231–235. https://doi.org/10.1016/j.bbrc.2009.04.093
Comin-Anduix B, Escuin-Ordinas H, Ibarrondo FJ (2016) Tremelimumab: research and clinical development. Onco Targets Ther 9:1767–1776. https://doi.org/10.2147/OTT.S65802
Cortés M, Sanchez-Moral L, de Barrios O et al (2017) Tumor-associated macrophages (TAMs) depend on ZEB1 for their cancer-promoting roles. EMBO J 36(22):3336–3355. https://doi.org/10.15252/embj.201797345
Das M, Zhu C, Kuchroo VK (2017) Tim-3 and its role in regulating anti-tumor immunity. Immunol Rev 276(1):97–111. https://doi.org/10.1111/imr.12520
De Araújo FV, Carrillo-Gálvez AB, Martín F, Anderson P (2018) TGF-β and mesenchymal stromal cells in regenerative medicine, autoimmunity and cancer. Cytokine Growth Factor Rev 43:25–37. https://doi.org/10.1016/j.cytogfr.2018.06.002
Digklia A, Wagner AD (2016) Advanced gastric cancer: current treatment landscape and future perspectives. World J Gastroenterol 22(8):2403–2414. https://doi.org/10.1016/j.ejca.2015.07.026
Du W, Liu N, Zhang Y et al (2020) PLOD2 promotes aerobic glycolysis and cell progression in colorectal cancer by upregulating HK2. Biochem Cell Biol 98(3):386–395. https://doi.org/10.1139/bcb-2019-0256
Essner JJ, Chen E, Ekker SC (2006) Syndecan-2. Int J Biochem Cell Biol 38(2):152–156. https://doi.org/10.1016/j.biocel.2005.08.012
Hamilton G, Rath B (2017) Avelumab: combining immune checkpoint inhibition and antibody-dependent cytotoxicity. Expert Opin Biol Ther 17(4):515–523. https://doi.org/10.1080/14712598.2017.1294156
Han I, Park H, Oh ES (2004) New insights into syndecan-2 expression and tumourigenic activity in colon carcinoma cells. J Mol Histol 35(3):319–326. https://doi.org/10.1023/b:hijo.0000032363.78829.4e
Han J, Choi YL, Kim H et al (2017) MMP11 and CD2 as novel prognostic factors in hormone receptor-negative, HER2-positive breast cancer. Breast Cancer Res Treat 164(1):41–56. https://doi.org/10.1007/s10549-017-4234-4
Henderson NC, Arnold TD, Katamura Y et al (2013) Targeting of αv integrin identifies a core molecular pathway that regulates fibrosis in several organs. Nat Med 19(12):1617–1624. https://doi.org/10.1038/nm.3282
Higuchi M, Kato T, Yoshida S et al (2015) PRRX1- and PRRX2-positive mesenchymal stem/progenitor cells are involved in vasculogenesis during rat embryonic pituitary development. Cell Tissue Res 361(22):557–565. https://doi.org/10.1007/s00441-015-2128-5
Hua R, Yu J, Yan X et al (2020) Syndecan-2 in colorectal cancer plays oncogenic role via epithelial-mesenchymal transition and MAPK pathway. Biomed Pharmacother 121:109630. https://doi.org/10.1016/j.biopha.2019.109630
Jia YY, Yu Y, Li HJ (2021) POSTN promotes proliferation and epithelial-mesenchymal transition in renal cell carcinoma through ILK/AKT/mTOR pathway. J Cancer 12(14):4183–4195. https://doi.org/10.7150/jca.51253
Jiang H, Yu D, Yang P et al (2022) Revealing the transcriptional heterogeneity of organ-specific metastasis in human gastric cancer using single-cell RNA sequencing. Clin Transl Med 12(2):e730. https://doi.org/10.1002/ctm2.730
Karimi P, Islami F, Anandasabapathy S, Freedman ND, Kamangar F et al (2014) Gastric cancer: descriptive epidemiology, risk factors, screening, and prevention. Cancer Epidemiol Biomark Prev 23(5):700–713. https://doi.org/10.1158/1055-9965.EPI-13-1057
Kemper M, Schiecke A, Maar H et al (2021) Integrin alpha-V is an important driver in pancreatic adenocarcinoma progression. J Exp Clin Cancer Res 40(1):214. https://doi.org/10.1186/s13046-021-01946-2
Khan Z, Marshall JF (2016) The role of integrins in TGFβ activation in the tumour stroma. Cell Tissue Res 365(3):657–673. https://doi.org/10.1007/s00441-016-2474-y
Kim N, Nam YS, Im KI et al (2015) IL-21-expressing mesenchymal stem cells prevent lethal B-cell lymphoma through efficient delivery of IL-21, which redirects the immune system to target the tumor. Stem Cells Dev 24(23):2808–2821. https://doi.org/10.1089/scd.2015.0103
Kou YB, Zhang SY, Zhao BL et al (2013) Knockdown of MMP11 inhibits proliferation and invasion of gastric cancer cells. Int J Immunopathol Pharmacol 26(2):361–370. https://doi.org/10.1177/039463201302600209
Lei Y, Tang R, Xu J et al (2021) Applications of single-cell sequencing in cancer research: progress and perspectives. J Hematol Oncol 14(1):91. https://doi.org/10.1186/s13045-021-01105-2
Li W, Zhang X, Wu F et al (2019) Gastric cancer-derived mesenchymal stromal cells trigger M2 macrophage polarization that promotes metastasis and EMT in gastric cancer. Cell Death Dis 10(12):918. https://doi.org/10.1038/s41419-019-2131-y
Liu T, Xia R, Li C et al (2021) mRNA expression level of CDH2, LEP, POSTN, TIMP1 and VEGFC modulates 5-fluorouracil resistance in colon cancer cells. Exp Ther Med 22(3):1023. https://doi.org/10.3892/etm.2021.10455
Loeser H, Scholz M, Fuchs H et al (2020) Integrin alpha V (ITGAV) expression in esophageal adenocarcinoma is associated with shortened overall-survival. Sci Rep 10(1):18411. https://doi.org/10.1038/s41598-020-75085-7
Lu J, Chen Y, Zhang X et al (2022) A novel prognostic model based on single-cell RNA sequencing data for hepatocellular carcinoma. Cancer Cell Int 22(1):8. https://doi.org/10.1186/s12935-022-02469-2
Lv FJ, Tuan RS, Cheung KM, Leung VY (2014) Concise review: the surface markers and identity of human mesenchymal stem cells. Stem Cells 32(6):1408–1419. https://doi.org/10.1002/stem.1681
Malanchi I, Santamaria-Martínez A, Susanto E et al (2011) Interactions between cancer stem cells and their niche govern metastatic colonization. Nature 481(7379):85–89. https://doi.org/10.1038/nature10694
Mangiola S, Doyle MA, Papenfuss AT (2021) Interfacing seurat with the R tidy universe. Bioinformatics 37(22):4100–4107. https://doi.org/10.1093/bioinformatics/btab404
Marin JJ, Al-Abdulla R, Lozano E et al (2016) Mechanisms of resistance to chemotherapy in gastric cancer. Anticancer Agents Med Chem 16(3):318–334. https://doi.org/10.2174/1871520615666150803125121
Marini I, Siegemund M, Hutt M, Kontermann RE, Pfizenmaier K (2017) Antitumor activity of a mesenchymal stem cell line stably secreting a tumor-targeted TNF-related apoptosis-inducing ligand fusion protein. Front Immunol 8:536. https://doi.org/10.3389/fimmu.2017.00536
Miller JD, Lankford SM, Adler KB, Brody AR (2010) Mesenchymal stem cells require MARCKS protein for directed chemotaxis in vitro. Am J Respir Cell Mol Biol 43(3):253–258. https://doi.org/10.1165/rcmb.2010-0015RC
Mirzaei H, Ahebkar A, Avan A et al (2016) Application of mesenchymal stem cells in melanoma: a potential therapeutic strategy for delivery of targeted agents. Curr Med Chem 23(5):455–463. https://doi.org/10.2174/0929867323666151217122033
Naito T, Yuge R, Kitadai Y et al (2021) Mesenchymal stem cells induce tumor stroma formation and epithelial–mesenchymal transition through SPARC expression in colorectal cancer. Oncol Rep 45(6):104. https://doi.org/10.3892/or.2021.8055
Nakayama Y, Mimura K, Kua LF et al (2020) Immune suppression caused by PD-L2 expression on tumor cells in gastric cancer. Gastric Cancer 23(6):961–973. https://doi.org/10.1007/s10120-020-01079-z
Newman AM, Liu CL, Green MR et al (2015) Robust enumeration of cell subsets from tissue expression profiles. Nat Methods 12(5):453–457. https://doi.org/10.1038/nmeth.3337
Ni Z, Xing D, Zhang T et al (2021) Tumor-infiltrating B cell is associated with the control of progression of gastric cancer. Immunol Res 69(1):43–52. https://doi.org/10.1007/s12026-020-09167-z
Ralph C, Elkord E, Burt DJ et al (2010) Modulation of lymphocyte regulation for cancer therapy: a phase II trial of tremelimumab in advanced gastric and esophageal adenocarcinoma. Clin Cancer Res 16(5):1662–1672. https://doi.org/10.1158/1078-0432.CCR-09-2870
Sheng X, Li Y, Li Y et al (2019) PLOD2 contributes to drug resistance in laryngeal cancer by promoting cancer stem cell-like characteristics. BMC Cancer 19(1):840. https://doi.org/10.1186/s12885-019-6029-y
Shitara K, Van Cutsem E, Bang YJ et al (2020) Efficacy and safety of pembrolizumab or pembrolizumab plus chemotherapy vs chemotherapy alone for patients with first-line, advanced gastric cancer: the KEYNOTE-062 phase 3 randomized clinical trial. JAMA Oncol 6(10):1571–1580. https://doi.org/10.1001/jamaoncol.2020.3370
Song P, Li W, Wu X et al (2022) Integrated analysis of single-cell and bulk RNA-sequencing identifies a signature based on B cell marker genes to predict prognosis and immunotherapy response in lung adenocarcinoma. Cancer Immunol Immunother 71(10):2341–2354. https://doi.org/10.1007/s00262-022-03143-2
St Paul M, Ohashi PS (2020) The roles of CD8(+) T cell subsets in antitumor immunity. Trends in Cell Biol 30(9):695–704. https://doi.org/10.1016/j.tcb.2020.06.003
Sun XJ, Liu S, Wang J et al (2018) Annexin A5 regulates hepatocarcinoma malignancy via CRKI/II-DOCK180-RAC1 integrin and MEK-ERK pathways. Cell Death Dis 9(6):637. https://doi.org/10.1038/s41419-018-0685-8
Sun G, Li Z, Rong D et al (2021) Single-cell RNA sequencing in cancer: applications, advances, and emerging challenges. Mol Therapy Oncolytics 21:183–206. https://doi.org/10.1016/j.omto.2021.04.001
Sung H, Ferlay J, Siegel RL et al (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. Cancer J Clin 71(3):209–249. https://doi.org/10.3322/caac.21660
Takei S, Kawazoe A, Shitara K (2022) The New Era of Immunotherapy in Gastric Cancer. Cancers (basel) 14(4):1054. https://doi.org/10.3390/cancers14041054
Ullah I, Subbarao RB, Rho GJ (2015) Human mesenchymal stem cells - current trends and future prospective. Biosci Rep 35(2):e191. https://doi.org/10.1042/BSR20150025
Verlato G, Marrelli D, Accordini S et al (2015) Short-term and long-term risk factors in gastric cancer. World J Gastroenterol 21(21):6434–6443. https://doi.org/10.3748/wjg.v21.i21.6434
Wang X, Guo J, Dai M et al (2020) PLOD2 increases resistance of gastric cancer cells to 5-fluorouracil by upregulating BCRP and inhibiting apoptosis. J Cancer 11(2):3467–3475. https://doi.org/10.7150/jca.41828
Wei E, Reisinger A, Li J et al (2022) Integration of scRNA-Seq and TCGA RNA-Seq to analyze the heterogeneity of HPV+ and HPV-cervical cancer immune cells and establish molecular risk models. Front Oncol 12:860900. https://doi.org/10.3389/fonc.2022.860900
Woodward A, Faria GNF, Harrison RG (2022) Annexin A5 as a targeting agent for cancer treatment. Cancer Lett 547:215857. https://doi.org/10.1016/j.canlet.2022.215857
Wu E, Mari BP, Wang F et al (2001) Stromelysin-3 suppresses tumor cell apoptosis in a murine model. J Cell Biochem 82(4):549–555. https://doi.org/10.1002/jcb.1181
Wu XB, Liu Y, Wang GH et al (2016) Mesenchymal stem cells promote colorectal cancer progression through AMPK/mTOR-mediated NF-κB activation. Sci Rep 6:21420. https://doi.org/10.1038/srep21420
Wu S, Wang Y, Yuan Z et al (2018) Human adipose-derived mesenchymal stem cells promote breast cancer MCF7 cell epithelial–mesenchymal transition by cross interacting with the TGF-β/Smad and PI3K/AKT signaling pathways. Mol Med Rep 19(1):177–186. https://doi.org/10.3892/mmr.2018.9664
Xue G, Hao LQ, Ding FX et al (2009) Expression of annexin a5 is associated with higher tumor stage and poor prognosis in colorectal adenocarcinomas. J Clin Gastroenterol 43(9):831–837. https://doi.org/10.1097/MCG.0b013e31819cc731
Yan Y, Zhang J, Li JH et al (2016) High tumor-associated macrophages infiltration is associated with poor prognosis and may contribute to the phenomenon of epithelial-mesenchymal transition in gastric cancer. Onco Targets Ther 9:3975–3983. https://doi.org/10.2147/OTT.S103112
Yang W, Soares J, Greninger P et al (2013) Genomics of drug sensitivity in cancer (GDSC): a resource for therapeutic biomarker discovery in cancer cells. Nucleic Acids Res 41(D1):D955-961. https://doi.org/10.1093/nar/gks1111
Yang J, Zhan XZ, Malola J et al (2020) The multiple roles of Thy-1 in cell differentiation and regeneration. Differentiation 113:38–48. https://doi.org/10.1016/j.diff.2020.03.003
Zhang X, Lan Y, Xu J et al (2019) Cell marker: a manually curated resource of cell markers in human and mouse. Nucleic Acids Res 47(D1):D721–D728. https://doi.org/10.1093/nar
Zhou WT, Jin WL (2021) B7–H3/CD276: an emerging cancer immunotherapy. Front Immunol 12:701006. https://doi.org/10.3389/fimmu.2021.701006
Zhu W, Huang L, Li Y et al (2012) Exosomes derived from human bone marrow mesenchymal stem cells promote tumor growth in vivo. Cancer Lett 315(1):28–37. https://doi.org/10.1016/j.canlet.2011.10.002
Ziemba BP, Burke JE, Masson G, Williams RL, Falke JJ (2016) Regulation of PI3K by PKC and MARCKS: single-molecule analysis of a reconstituted signaling pathway. Biophys J 110(8):1811–1825. https://doi.org/10.1016/j.bpj.2016.03.001
Funding
This study was supported by the Pang Dexiang Famous Veteran Chinese Medicine Specialist Inheritance Studio (2A12012014) and the Zhejiang Provincial Traditional Chinese Medicine Science and Technology Programme Projects (2021ZB128).
Author information
Authors and Affiliations
Contributions
GWC and SKY designed the research, SKY and CBY wrote the manuscript, SKY and CBY performed the in vitro experiments, GWC and SKY analyzed and interpreted the data. All authors contributed to this work.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
No ethics approval and informed consent are needed in the current work.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Shen, K., Chen, B. & Gao, W. Integrated single-cell RNA sequencing analysis reveals a mesenchymal stem cell-associated signature for estimating prognosis and drug sensitivity in gastric cancer. J Cancer Res Clin Oncol 149, 11829–11847 (2023). https://doi.org/10.1007/s00432-023-05058-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-023-05058-6