Validity and prognostic significance of sperm protein 17 as a tumor biomarker for epithelial ovarian cancer: a retrospective study.

BACKGROUND: Prior small studies have shown increased expression of sperm protein 17 (Sp17) in epithelial ovarian cancer (EOC) tissue and suggest Sp17 as a potential biomarker for EOC. However, how Sp17 expression varies with histology, grade, and stage of EOC and its expression in other ovarian neoplasms has not been defined. It is unknown whether patients with EOC have elevated serum Sp17 levels or if Sp17 expression is associated with survival outcomes. METHODS: The study included 982 patients with benign, borderline, and malignant ovarian neoplasms and normal ovary. There were 878 patients with tissue only, 39 with serum only, and 65 with matching serum and tissue. Immunohistochemical (IHC) staining with anti-Sp17 antibody was performed on tissue specimens and the intensity scored as weak, moderate, or strong. A sandwich enzyme-linked immunosorbent assay (ELISA) was performed to measure Sp17 sera concentrations. RESULTS: Sp17 expression was most commonly seen in serous cystadenomas (83%) and serous borderline tumors (100%). Of the 773 EOC specimens, 223 (30%) expressed Sp17. Grade and histology were significantly associated with Sp17 expression among EOC specimens (p < 0.001) on both univariate and multivariable analysis, with grade 1 serous adenocarcinomas showing the highest expression (51%). Sp17 expression was limited in other benign and non-epithelial malignant neoplasms. Neither Sp17 tissue expression nor serum concentration correlated with survival outcomes. Serum concentrations were higher in patients with Sp17 tissue expression, and the highest concentrations were noted among patients with serous and clear cell adenocarcinomas. CONCLUSIONS: Sp17 is highly expressed in benign, borderline, and low grade malignant serous ovarian neoplasms and can be quantified in serum. Sp17 expression may have diagnostic significance in this subset of patients.

Umbilical cord blood-derived dendritic cells infected by adenovirus for SP17 expression induce antigen-specific cytotoxic T cells against NSCLC cells.

Sperm protein 17 (SP17), a cancer/testis antigen, is expressed by non-small cell lung cancer (NSCLC). This study examined whether dendritic cells (DC) from human umbilical cord blood (UCB) could be induced for SP17 expression and induce antigen-specific CD8(+) cytotoxic T lymphocytes (CTLs) against NSCLC in vitro. We generated recombinant adenovirus of Ad-SP17 and control Ad-null. Infection with Ad-SP17, but not control, induced higher levels of SP17 expression in UCB-derived DC-Ad-SP17. Infection with Ad-SP17 significantly increased the frequency of CD80(+), CD83(+), CD86(+), and HLA-DR(+) DC that produced higher levels of IL-12, but lower IL-10. Co-culture of DC-Ad-SP17 with autologous UCB lymphocytes induced high frequency of IFNγ(+) CD8(+) CTLs, which had selective cytotoxicity against SP17(+) lung cancer CRL-5922 cells in a HLA-I restrictive manner. Thus, UCB-derived DC modulated for SP17 expression induced antigen-specific anti-tumor immunity against SP17(+) NSCLC, and SP17 may be a valuable target for development of immunotherapy against SP17(+) NSCLC.

Sperm protein 17 and AKAP-associated sperm protein cancer/testis antigens are expressed in ciliated hepatic foregut cysts.

AIMS: Ciliated hepatic foregut cysts (CHFCs) are retained benign lesions of the liver. However, a case of squamous cell metaplasia and five cases of squamous cell carcinoma arising from a CHFC have been described. The potential of malignant transformation makes the identification of new biomarkers necessary. As the cancer/testis antigen sperm protein 17 (Sp17) has been detected in oral and oesophageal squamous cell carcinomas, the aim of this study was to investigate the expression of Sp17 and AKAP-associated sperm protein (ASP), which has a shared N-terminal sequence with Sp17, in four surgically resected CHFCs. METHODS AND RESULTS: CHFC specimens were taken from two patients who attended the Medical College of Wisconsin, Milwaukee, USA and two patients who attended the Fundación Jiménez Díaz, Madrid, Spain. CHFCs were found to be immunopositive for Sp17 and ASP. Both proteins were localized to the cytoplasm of ciliated cells lining the cysts, and their cilia. Confocal microscopy demonstrated that Sp17 and ASP overlapped in the same region of the cell. CONCLUSION: Sp17 and ASP cancer/testis antigens were found in ciliated cells of four CHFCs. Further characterization of Sp17 and ASP in patients with CHFCs may provide significant clues for understanding the molecular mechanisms underlying their predisposition to develop squamous cell carcinomas.

Enhanced Electrochemical Biosensing of the Sp17 Cancer Biomarker in Serum Samples via Engineered Two-Dimensional MoS2 Nanosheets on the Reduced Graphene Oxide Interface.

In the present investigation, we reported a label-free and highly effective immunosensor for the first time employing a nanostructured molybdenum disulfide nanosheets@reduced graphene oxide (nMoS2 NS@rGO) nanohybrid interface for the determination of sperm protein 17 (Sp17), an emerging cancer biomarker. We synthesized the nMoS2 NS@rGO nanohybrid using a one-step hydrothermal technique and then functionalized it with 3-aminopropyltriethoxysilane (APTES). Furthermore, the anti-Sp17 monoclonal antibodies were covalently attached to the APTES/nMoS2 NS@rGO/indium tin oxide (ITO) electrode utilizing 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-N-hydroxy succinimide (EDC-NHS) coupling chemistry. Bovine serum albumin (BSA) was then used to block nonspecific binding regions on the anti-Sp17/APTES/nMoS2 NS@rGO/ITO bioelectrode. The morphological and structural features of the synthesized nanohybrid and the modified electrodes were studied using transmission electron microscopy, scanning electron microscopy with energy dispersive X-ray (EDX) composition studies, atomic force microscopy, Fourier transform infrared spectroscopy, and Raman spectroscopy. The immunoreaction between the Sp17 antigen and anti-Sp17 antibodies on the surface of the BSA/anti-Sp17/APTES/nMoS2 NS@rGO/ITO sensing bioelectrode was applied as the basis for the detection technique, which measured the electrocatalytic current and impedimetric response change. The designed BSA/anti-Sp17/APTES/nMoS2 NS@rGO/ITO bioelectrode showed improved amperometric and impedimetric biosensing performance in the response studies, including remarkable sensitivity (23.2 µA ng-1mL cm-2 and 0.48 kΩ mL ng-1 cm-2), wider linearity (0.05-8 and 1-8 ng mL-1), an excellent lower detection limit (0.13 and 0.23 ng mL-1), and a rapid response time of 20 min. The biosensor exhibited impressive storage durability lasting 7 weeks and showed remarkable precision in identifying Sp17 in serum samples from cancer patients, as confirmed using the enzyme-linked immunosorbent assay method.

Biocompatible epoxysilane substituted polymer-based nano biosensing platform for label-free detection of cancer biomarker SP17 in patient serum samples.

Herein, we report the results of the studies relating to developing a simple, sensitive, cost-effective, and disposable electrochemical-based label-free immunosensor for real-time detection of a new cancer biomarker, sperm protein-17 (SP17), in complex serum samples. An indium tin oxide (ITO) coated glass substrate modified with self-assembled monolayers (SAMs) of 3-glycidoxypropyltrimethoxysilane (GPTMS) was functionalized via covalent immobilization of monoclonal anti-SP17 antibodies using EDC(1-(3-(dimethylamine)-propyl)-3-ethylcarbodiimide hydrochloride) - NHS (N-hydroxy succinimide) chemistry. The developed immunosensor platform (BSA/anti-SP17/GPTMS@SAMs/ITO) was characterized via scanning electron microscopy (SEM), atomic force microscopy (AFM), contact angle (CA), Fourier transform infrared (FT-IR) spectroscopic, and electrochemical techniques such as cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) techniques. The fabricated BSA/anti-SP17/GPTMS@SAMs/ITO immunoelectrode platform was used to measure changes in the magnitude of the current of the electrodes through an electrochemical CV and DPV technique. A calibration curve between current and SP17 concentrations exhibited a broad linear detection range of (100-6000 & 50-5500 pg mL-1), with enhanced sensitivity (0.047 & 0.024 µA pg mL-1 cm-2), limit of detection (LOD) and limit of quantification (LOQ) of 47.57 & 142.9 pg mL-1 and 158.58 & 476.3 pg mL-1, by CV and DPV technique, respectively with a rapid response time of 15 min. It possessed exceptional repeatability, outstanding reproducibility, five-time reusability, and high stability. The biosensor's performance was evaluated in human serum samples, giving satisfactory findings obtained via the commercially available enzyme-linked immunosorbent assay (ELISA) technique, proving the clinical applicability for early diagnosis of cancer patients. Moreover, various in vitro studies in murine fibroblast cell line L929 have been performed to assess the cytotoxicity of GPTMS. The results demonstrated that GPTMS has excellent biocompatibility and can be used for biosensor fabrication.

Fabrication of alkoxysilane substituted polymer-modified disposable biosensing platform: Towards sperm protein 17 sensing as a new cancer biomarker.

SP17 is a mammalian protein found in the testis and spermatozoa that have been identified as a tumor-associated antigen in a range of human cancers. A unique method for fabricating the first ultrasensitive, selective, and label-free immunosensor for the detection of SP17, a new cancer biomarker in complicated serum samples, is presented in this paper. This immunosensor was also the first biosensor built using a disposable ITO sheet modified with an aminosilane known as APTMS as an immobilization platform for fabricating the SP17 biosensor. The immobilization of chemical and biological species onto the electrode surface was cross-verified by various analytical and morphological techniques. Stepwise modifications done on the immunoelectrodes were also studied using electrochemical techniques. Selective interaction between anti-SP17 and SP17 with varying concentrations (100-5000 pg mL-1) was measured with the DPV technique. The immunosensor exhibited low LOD and LOQ of 70.07 and 233.57 pg mL-1, respectively, with a sensitivity of 0.013 µA mL pg-1 cm-2. The fabricated immunosensor performance was analyzed by quantifying the SP17 concentrations in patient serum samples. The data obtained from the developed immunosensor demonstrated excellent reproducibility, repeatability, and selectivity among various interferants, including cancer biomarkers. Further, the observed results have been validated via ELISA, which showed good agreement with the electrochemical results. This could establish a new platform for detecting other cancer biomarkers and can be employed for clinical diagnostics applications.

Sperm protein 17 targeting for epithelialovarian cancer treatment in the eraof modern immunoengineering.

Due to the vague symptomatology of the disease and a lack of effective screening methods, most patients with epithelial ovarian cancer (EOC) present late in their disease. Despite advances in chemotherapeutic agents, the prognosis of these patients has uniformly been extremely poor. Although cisplatin-based chemotherapy regimens induce responses in most of these patients, the patients invariably experience disease progression or relapses. In an attempt to improve the treatment outcome using a different therapeutic approach, immunotherapy was investigated nearly 20 years ago. Many tumor antigens that are potentially suitable for specific immunotherapy were identified, and many immunotherapeutic approaches were attempted. However, although some responses were observed, the results from clinical studies were generally disappointing. Recent advances in immunoengineering and successes observed among patients treated for refractory/relapsed hematologic malignancies have rekindled the interest to revisit specific cellular immunotherapy in EOC. In this review, we provide the rationale for immunotherapy of EOC, discuss the results of some of the historical studies on the use of cellular immunotherapy in EOC, outline the principles of modern immunoengineering that could be applied to treat the disease, and propose the re-evaluation of the cancer-testis antigen, Sperm protein 17, for targeting by using modern immunoengineering technology.

Sperm Protein 17 Expression by Murine Epithelial Ovarian Cancer Cells and Its Impact on Tumor Progression.

The cancer testis antigen sperm protein 17 (Sp17) is a promising antigenic target in epithelial ovarian cancer (EOC) vaccine development. However, its role in ovarian cancer is unclear. We isolated and expanded Sp17⁺ and Sp17- clones from the murine EOC cell line ID8, and compared their in-vitro cell growth characteristics and in-vivo tumorigenicity. We also examined the potential co-expression of molecules that may influence cancer cell survival and interaction with immune cells. These include stimulatory and immunosuppressive molecules, such as major histocompatibility class I molecules (MHC I), MHC II, cytotoxic T lymphocyte associated antigen-4 (CTLA-4), CD73, CD39, tumor necrosis factor receptor II (TNFRII), signal transducer and activator of transcription 3 (STAT3) and programmed death-ligand 1 (PD-L1). Whilst the presence of Sp17 was not correlated with the ID8 cell proliferation/growth capacity in vitro, it was critical to enable progressive tumor formation in vivo. Flow cytometry revealed that Sp17⁺ ID8 cells displayed higher expression of both STAT3 and PD-L1, whilst MHC II expression was lower. Moreover, Sp17high (PD-L1⁺MHCII-) cell populations showed significantly enhanced resistance to Paclitaxel-induced cell death in vitro compared to Sp17low (PD-L1-MHCII⁺) cells, which was associated in turn with increased STAT3 expression. Together, the data support Sp17 as a factor associated with in-vivo tumor progression and chemo-resistance, validating it as a suitable target for vaccine development.

Mapping T and B cell epitopes in sperm protein 17 to support the development of an ovarian cancer vaccine.

Ovarian cancer (OC) is the seventh most common cancer in women worldwide, and the leading cause of death from gynaecological malignancy. Immunotherapeutic strategies including cancer vaccines are considered less toxic and more specific than current treatments. Sperm surface protein (Sp17) is a protein aberrantly expressed in primary as well as in metastatic lesions in >83% of ovarian cancer patients. Vaccines based on the Sp17 protein are immunogenic and protective in animal models. To map the immunogenic regions and support the development of human Sp17 peptide based vaccines, we used 6 overlapping peptides of the human Sp17 sequence adjuvanted with CpG to immunise humanised HLA-A2.1 transgenic C57BL/6 mice, and assessed immunogenicity by ELISPOT and ELISA. No CD8 T cells were found to be induced to a comprehensive panel of 10 HLA-A2.1 or H-2K(b) binding predicted epitopes. However, one of the 6 peptides, hSp17111-142, induced high levels of antibodies and IFN-γ producing T cells (but not IL-17 or IL-4) both in C57BL/6 and in C57BL/6-HLA-A2.1 transgenic mice. C57BL/6 mice immunised with CpG adjuvanted hSp17111-142 significantly prolonged the life-span of the mice bearing the ovarian carcinoma ID8 cell line. We further mapped the immuno-dominant B and T cell epitope regions within hSp17111-142 using ELISPOT and competition ELISA. Herein, we report the identification of a single immuno-dominant B cell (134-142 aa) epitope and 2 T helper 1 (Th1) cell epitopes (111-124 aa and 124-138 aa). These result together support further exploration of hSp17111-142 peptide formulations as vaccines against ovarian cancer.

Chimeric antigen receptor engineering: a right step in the evolution of adoptive cellular immunotherapy.

Cancer immunotherapy comprises different therapeutic strategies that exploit the use of distinct components of the immune system, with the common goal of specifically targeting and eradicating neoplastic cells. These varied approaches include the use of specific monoclonal antibodies, checkpoint inhibitors, cytokines, therapeutic cancer vaccines and cellular anticancer strategies such as activated dendritic cell (DC) vaccines, tumor-infiltrating lymphocytes (TILs) and, more recently, genetically engineered T cells. Each one of these approaches has demonstrated promise, but their generalized success has been hindered by the paucity of specific tumor targets resulting in suboptimal tumor responses and unpredictable toxicities. This review will concentrate on recent advances on the use of engineered T cells for adoptive cellular immunotherapy (ACI) in cancer.

Sperm protein 17 is an oncofetal antigen: a lesson from a murine model.

Sperm protein 17 (Sp17) was originally identified in the flagellum of spermatozoa and subsequently included in the subfamily of tumor-associated antigens known as cancer-testes antigens (CTA). Sp17 has been associated with the motility and migratory capacity in tumor cells, representing a link between gene expression patterns in germinal and tumor cells of different histological origins. Here we review the relevance of Sp17 expression in the mouse embryo and cancerous tissues, and present additional data demonstrating Sp17 complex expression pattern in this murine model. The expression of Sp17 in embryonic as well as adult neoplastic cells, but not normal tissues, suggests this protein should be considered an "oncofetal antigen." Further investigations are necessary to elucidate the mechanisms and functional significance of Sp17 aberrant expression in human adult cells and its implication in the pathobiology of cancer.