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A Key Regulator of Tumor Progression and Immunomodulation cancers Review Osteopontin: A Key Regulator of Tumor Progression and Immunomodulation Hannah R. Moorman 1, Dakota Poschel 1,2,3, John D. Klement 1,2,3, Chunwan Lu 1,2,3, Priscilla S. Redd 4 and Kebin Liu 1,2,3,* 1 Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, GA 30912, USA; [email protected] (H.R.M.); [email protected] (D.P.); [email protected] (J.D.K.); [email protected] (C.L.) 2 Georgia Cancer Center, Medical College of Georgia, Augusta, GA 30912, USA 3 Charlie Norwood VA Medical Center, Augusta, GA 30904, USA 4 Chemedimmune Inc., Augusta, GA 30912, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-706-721-9483 Received: 22 October 2020; Accepted: 10 November 2020; Published: 15 November 2020 Simple Summary: Anti-PD-1/PD-L1 and anti-CTLA-4-based immune checkpoint blockade (ICB) immunotherapy have recently emerged as a breakthrough in human cancer treatment. Durable efficacy has been achieved in many types of human cancers. However, not all human cancers respond to current ICB immunotherapy and only a fraction of the responsive cancers exhibit efficacy. Osteopontin (OPN) expression is highly elevated in human cancers and functions as a tumor promoter. Emerging data suggest that OPN may also regulate immune cell function in the tumor microenvironment. This review aims at OPN function in human cancer progression and new findings of OPN as a new immune checkpoint. We propose that OPN compensates PD-L1 function to promote tumor immune evasion, which may underlie human cancer non-response to current ICB immunotherapy. Abstract: OPN is a multifunctional phosphoglycoprotein expressed in a wide range of cells, including osteoclasts, osteoblasts, neurons, epithelial cells, T, B, NK, NK T, myeloid, and innate lymphoid cells. OPN plays an important role in diverse biological processes and is implicated in multiple diseases such as cardiovascular, diabetes, kidney, proinflammatory, fibrosis, nephrolithiasis, wound healing, and cancer. In cancer patients, overexpressed OPN is often detected in the tumor microenvironment and elevated serum OPN level is correlated with poor prognosis. Initially identified in activated T cells and termed as early T cell activation gene, OPN links innate cells to adaptive cells in immune response to infection and cancer. Recent single cell RNA sequencing revealed that OPN is primarily expressed in tumor cells and tumor-infiltrating myeloid cells in human cancer patients. Emerging experimental data reveal a key role of OPN is tumor immune evasion through regulating macrophage polarization, recruitment, and inhibition of T cell activation in the tumor microenvironment. Therefore, in addition to its well-established direct tumor cell promotion function, OPN also acts as an immune checkpoint to negatively regulate T cell activation. The OPN protein level is highly elevated in peripheral blood of human cancer patients. OPN blockade immunotherapy with OPN neutralization monoclonal antibodies (mAbs) thus represents an attractive approach in human cancer immunotherapy. Keywords: osteopontin; CD44; integrin; tumor-associated macrophage; MDSCs; immune evasion; immune checkpoint 1. Introduction Osteopontin (OPN), a phosphorylated glycoprotein, was originally identified as a secreted form in bone and later discovered as an intracellular protein [1,2]. OPN is widely expressed in osteoclasts; Cancers 2020, 12, 3379; doi:10.3390/cancers12113379 www.mdpi.com/journal/cancers Cancers 2020, 12 2 of 31 1. Introduction Cancers 2020, 12, 3379 2 of 31 Osteopontin (OPN), a phosphorylated glycoprotein, was originally identified as a secreted form in bone and later discovered as an intracellular protein [1,2]. OPN is widely expressed in osteoclasts; osteoblasts;osteoblasts; epithelial epithelial cells cells of of th thee breast, breast, kidney, kidney, and and skin; skin; nerve nerve cells; cells; vascular vascular smooth smooth muscle muscle cells; cells;endothelial endothelial cells; cells;and andfibroblasts fibroblasts [3,4]. [3 ,A4]. broa A broadd range range of ofimmune immune cells cells such such as as macrophages,macrophages, lymphocytes,lymphocytes, natural killer cells,cells, eosinophils, de dendriticndritic cells, and microglia express OPN [5–11]. [5–11]. GivenGiven OPN’sOPN’s widewide expressionexpression profile,profile, itit followsfollows thatthat OPNOPN isis notnot onlyonly importantimportant forfor normalnormal biologicalbiological functioningfunctioning suchsuch as as bone bone remodeling, remodeling, immunity, immunity, and inflammation, and inflammation, but also playsbut also a role plays in pathological a role in processespathological such processes as liver fibrosis,such as liver atherosclerosis, fibrosis, atheroscle and cancerrosis, [12 and,13]. cancer [12,13]. InIn cancercancer patients,patients, OPNOPN isis oftenoften overexpressedoverexpressed inin cellscells ofof thethe tumortumor microenvironmentmicroenvironment andand elevatedelevated in the peripheral blood. blood. OPN OPN overexpressi overexpressionon has has been been associated associated with with worse worse prognosis prognosis in inmany many cancers cancers including including glioblastoma glioblastoma multiforme, multiforme, hepatocellular hepatocellular carcinoma, carcinoma, colorectal colorectal cancer, cancer, lung lungcancer, cancer, breast breast cancer, cancer, bladder bladder cancer, cancer, melanoma, melanoma, head headand neck and necksquamous squamous cell carcinoma, cell carcinoma, and acute and acutemyeloid myeloid leukemia leukemia [14–39]. [14 –OPN39]. OPNcontributes contributes to the to malignancy the malignancy of cancer of cancer through through the the promotion promotion of ofmetastasis, metastasis, maintenance maintenance of ofa astem-like stem-like phenotyp phenotype,e, epithelial epithelial to to mesenchymal transformation,transformation, activationactivation ofof cell proliferation pathways, chemotherapeutic andand radiation resistance, andand interference withwith immuneimmune functioning.functioning. As such,such, understanding OPN’sOPN’s regulationregulation andand mechanismsmechanisms ofof actionaction willwill provideprovide potentialpotential treatmenttreatment optionsoptions inin thethe future.future. In this review, we attempt to address the rolerole ofof OPNOPN inin cancerscancers includingincluding clinicalclinical data,data, mechanisms,mechanisms, regulation,regulation, andand potentialpotential treatmenttreatmentstrategies. strategies. 2.2. The OPN Protein OPNOPN isis aa membermember ofof thethe SIBLINGSIBLING (Small(Small Integrin-BindingIntegrin-Binding Ligand,Ligand, N-linkedN-linked Glycoprotein)Glycoprotein) familyfamily ofof proteinsproteins [[40].40]. OPN, also known as secreted phosphoprotein 1,1, isis encoded byby a single-copy gene,gene, spp1spp1, ,on on human human chromosome chromosome 4 near 4 near the centromere the centromere [41]. It [41 has]. several It has conserved several conserved structural structuraldomains including domains an including RGD domain an RGD (arginine-glycine-aspartate), domain (arginine-glycine-aspartate), SVVYGLR domain SVVYGLR (Serine-valine- domain (Serine-valine-valine-tyrosine-glutamate-leucine-arginine),valine-tyrosine-glutamate-leucine-arginine), heparin-binding heparin-binding domain, calcium-binding domain, calcium-binding domains, domains,cleavage sites cleavage for matrix sitesfor metalloproteinases, matrix metalloproteinases, and CD44 andreceptor-binding CD44 receptor-binding domain [42–49]. domain Five [42 OPN–49]. Fivesplice OPN variants splice have variants been identified. have been OPNa identified. (full-length OPNa (full-lengthisoform) contains isoform) all seven contains exons all sevenwhile OPNb exons whilelacks exon OPNb 5 lacksand OPNc exon 5lacks and exon OPNc 4 lacks(Figure exon 1). 4OPN4 (Figure lacks1). OPN4exons lacks4 and exons 5 and 4 OPN5 and 5 andcontains OPN5 all containsseven exons all seven plus exonsan extra plus exon an extra (yields exon alternativ (yields alternativee translation translation start) between start) between the third the and third fourth and fourthexons [50]. exons It [remains50]. It remains to be determined to be determined whether whether OPN4 and OPN4 OPN5 and transcripts OPN5 transcripts are translated are translated to protein. to protein.Unique Uniquephosphorylation phosphorylation sites and sites transglutaminase and transglutaminase cross-linking cross-linking sites allow sites allowfor structural for structural and andfunctional functional differences differences between between isoforms isoforms [48]. [ 48Differe]. Diffntialerential expression expression of OPN of OPN isoforms isoforms is likely is likely to tocontribute contribute to tocancer-specific cancer-specific progression. progression. For For example, example, OPNc OPNc supports supports anchorage anchorage independence independence and andinvasiveness invasiveness in breast in breast cancer cancer while OPNa while and OPNa OPNb and promote OPNb promotemigration migration in hepatocellular in hepatocellular carcinoma carcinoma[51–53]. [51–53]. FigureFigure 1.1. Human OPN protein structures.structures. The human SPP1 genes that encodes thethe OPNOPN proteinprotein hashas sevenseven exons.exons. Three alternativealternative splicedspliced OPNOPN variantsvariants translatetranslate toto protein.protein. TheThe receptor-interactingreceptor-interacting domainsdomains areare shown.shown. The respective receptorsreceptors forfor thesethese domainsdomains areare shownshown atat thethe bottom.bottom. Cancers
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