A. Sevko: A Portfolio of Translational Research Publications

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With significant experience in immune cell phenotyping and functional status assessment, as well as protein profiling, I have contributed to advancing our understanding of immune responses and their applications in therapeutic contexts. My work, detailed in several peer-reviewed publications, reflects a commitment to enhancing translational research in immuno-oncology, immune regulation, and biomarker discovery.

• Designing panels and analysis strategy for immune cell phenotyping using flow cytometry and other advanced techniques.
• Protein profiling design and analysis to identify biomarker signatures in oncology research.
• Collaborative work with multidisciplinary teams to generate actionable insights for drug development.

Selected publications  

1. Integrative Analysis of Immune and Protein Biomarkers in Urologic Oncology
   Urine biomarkers are crucial for monitoring patient responses in treating urological pathologies. Yet, analysing urine biomarkers poses several challenges, including ensuring specimen stability during transportation and analytical processing. This prospective feasibility study  demonstrates the compatibility of refrigerated urine shipment from the collection sites to analytical laboratories with both immunophenotyping and proteomic analysis and establishes clear logistical benefits for numerous clinical settings focused on monitoring patient immune responses in the urine matrix.  BMC Urol. 2024;24(1):276. 
2. Understanding systemic immune suppression in breast cancer: tumour-specific Tregs in the bone marrow On the basis of an analysis of the function, antigen specificity, and distribution of tumour antigen-reactive T cells and Tregs in patients with breast cancer and transgenic mouse tumour models, we showed that tumour-specific Tregs were selectively activated in the bone marrow (BM) and egressed into the peripheral blood.  Cancer Immunol Res. 2019;7(12):1998-2012. 
3. Results  of a pilot trial with Tadalafil in patients with metastatic melanoma (TaMe)  In the  open-label, dose de-escalation trial with tadalafil in pre-treated metastatic melanoma patients, we demonstrated that the stable patients displayed significantly higher numbers of CD8+ TIL in the centre of metastases before treatment as compared with progressive patients. Upon the therapy, they showed increased expression of ζ-chain (used as a marker of T cell activation) in CD8+ and CD4+TILs and CD8+T cells in the peripheral blood as compared with baseline.    Oncoimmunology. 2017;6(9):e1326440 
4. Immune Profiling Predicting Responses in ICI Melanoma Therapy
   In this study, we applied immune profiling techniques to identify immune signatures linked to Ipilimumab therapeutic efficacy.  We highlighted additional mechanisms of Ipilimumab effects and suggested levels of eosinophils, MDSCs, as well as related inflammatory factors S100A8/A9 and HMGB1 as novel complex predictive markers for patients who may benefit from the Ipilimumab therapy.  Clin Cancer Res. 2015;21(24):5453-9.  
5. Serum inflammatory mediators and circulating immunosuppressive cells as predictive biomarkers for  NSCLC radiofrequecy ablation response
   Here we analysed serum inflammatory factors as well as immunosuppressive cells in the peripheral blood to discover possible prognostic indicators. In the relapsing patients, we found early increase in TNF, CCL2 and CCL4, associated with increase in NO production by circulating MDSCs.  Clin Exp Immunol. 2015;180(3):467-74.  

• Understanding of local and systemic cancer-mediated immune changes in order to select targets for intervention.
• Developing  in vitro and in vivo models for investigation of cancer therapeutics mode of action in the immuno-oncology field.
• Translational research integrating preclinical and clinical data.
• Collaborative work with multidisciplinary teams to support IND submission, translation into clinical setting, and next-generation development.

 Selected publications

1. Coexpression of CD8a on the TCR-engineered CD4+ T cells
   T cells were transduced with a lentiviral vector to coexpress an affinity-enhanced HLA class I–restricted TCR directed against MAGE-A4 alongside a CD8α coreceptor.  Our results demonstrated enhanced specific anti-tumour CD4+ T-cell functions, which may potentially improve the depth and durability of antitumor responses in patients.    Journal of Immunotherapy 2023 46(4):p 132-144 .
2.  Functional tumour RNA transfer by tumour-derived extravesical vesicles in the genetic glioma and lung cancer models
   Using the engineered Cre recombinase-expressing glioma and carcinoma tumour cells which release Cre mRNA in various EV subfractions, including exosomes, in the animal models with a Cre reporter background leads to frequent recombination events at the tumour site. In both tumour models the majority of recombined cells were CD45+ leukocytes, predominantly Gr1+CD11b+ myeloid-derived suppressor cells (MDSCs).  Oncoimmunology. 2015;4(6):e1008371.      
3. Role of TRAIL signalling  in shaping tumour microenvironment: Cancer secretome
   We demonstrated that TRAIL-triggered cytokine secretion polarizes monocytes towards myeloid-derived suppressor cells (MDSCs) and tumour-associated macrophages both in in vitro models and in the lung cancer patients. TRAIL-R suppression in tumour cells impaired CCL2 production and diminished in vivo both lung MDSC presence and tumour growth.  Mol Cell 2017;65(4):730-742.e5. 
4. Temozolomide suppression of melanoma growth: effect on CCL2-driven signals
   The use of conventional agents in rationale combinations may result in optimization of therapy.  By showing that down-regulation of CCL2-driven signals by SAHA and temozolomide via JNK contributes to reduce melanoma growth, we provide a rationale for the therapeutic advantage of the drug combination. This combination strategy may be effective because of interference both with tumour cell and TME.  Oncotarget. 2014;5(12):4516-28. 
5. Myeloid cell immune suppression aggravated by  cyclophosphamide-mediated ablation of Tregs
   Cancer immune escape is frequently associated with the induction of an inappropriate immune response, i.e., a response that does not inhibit but can promote the tumour. Increased frequencies of tumour-infiltrating Tregs are associated with an impaired prognosis in several cancers. Thus, depletion of Tregs, e.g., by cyclophosphamide, was proposed as a means to boost immune responses to cancer. However, we provided evidence that cyclophosphamide exerted the unexpected effect of induction of myeloid-derived suppressor cells.   J Invest Dermatol 2013;133(6):1610-9. 
6. Paclitaxel in low doses mediates an anti-melanoma effect by inhibition of MDSCs and chronic inflammation
   In this study, we tested effects of ultralow noncytotoxic dose paclitaxel on functions of myeloid-derived suppressor cells (MDSCs), chronic inflammatory mediators, and T cell activities in the tumour microenvironment in vivo. Administration of paclitaxel significantly decreased accumulation and immunosuppressive activities of tumour-infiltrating MDSCs without alterations of the bone marrow haematopoiesis. This was associated with the inhibition of p38 MAPK activity, TNF-α and production, and S100A9 expression in MDSCs. The production of mediators of chronic inflammation in the tumour milieu also was diminished.   J Immunol 2013;190(5):2464-71.