Carolina Constantin

Protein bio-corona: critical issue in immune nanotoxicology.

Abstract With the expansion of the nanomedicine field, the knowledge focusing on the behavior of nanoparticles in the biological milieu has rapidly escalated. Upon introduction to a complex biological system, nanomaterials dynamically interact with all the encountered biomolecules and form the protein “bio-corona.” The decoration with these surface biomolecules endows nanoparticles with new properties. The present review will address updates of the protein bio-corona characteristics as influenced by nanoparticle’s physicochemical properties and by the particularities of the encountered biological milieu. Undeniably, bio-corona generation influences the efficacy of the nanodrug and guides the actions of innate and adaptive immunity. Exploiting the dynamic process of protein bio-corona development in combination with the new engineered horizons of drugs linked to nanoparticles could lead to innovative functional nanotherapies. Therefore, bio-medical nanotechnologies should focus on the interactions of nanoparticles with the immune system for both safety and efficacy reasons.

Chemically induced skin carcinogenesis: Updates in experimental models (Review)

Skin cancer is one of the most common malignancies affecting humans worldwide, and its incidence is rapidly increasing. The study of skin carcinogenesis is of major interest for both scientific research and clinical practice and the use of in vivo systems may facilitate the investigation of early alterations in the skin and of the mechanisms involved, and may also lead to the development of novel therapeutic strategies for skin cancer. This review outlines several aspects regarding the skin toxicity testing domain in mouse models of chemically induced skin carcinogenesis. There are important strain differences in view of the histological type, development and clinical evolution of the skin tumor, differences reported decades ago and confirmed by our hands-on experience. Using mouse models in preclinical testing is important due to the fact that, at the molecular level, common mechanisms with human cutaneous tumorigenesis are depicted. These animal models resemble human skin cancer development, in that genetic changes caused by carcinogens and pro-inflammatory cytokines, and simultaneous inflammation sustained by pro-inflammatory cytokines and chemokines favor tumor progression. Drugs and environmental conditions can be tested using these animal models. keeping in mind the differences between human and rodent skin physiology.

Immune-related biomarkers for diagnosis/prognosis and therapy monitoring of cutaneous melanoma

Skin melanoma, a life-threatening disease, has a recently reported worldwide increase in incidence, despite primary prevention. Skin melanoma statistics emphasize the need for finding markers related to the immune response of the host. The mechanisms that are able to over-power the local immune surveillance comprise molecules that can be valuable markers for diagnosis and prognosis. This article summarizes the immune markers that can monitor the disease stage and evaluate the efficacy of therapeutic interventions. Recent data regarding immunotherapy are presented in the context of tumor escape from immune surveillance and the immune molecules that are both targets and a means of monitoring. Perspectives for developing immune interventions for skin melanoma management and the position of tissue or soluble immune markers as a diagnostic/prognostic panel are evaluated. State-of-the-art technology is emphasized for developing immune molecular signatures for a complex characterization of the patient’s immunological status.

Immunomics in Skin Cancer - Improvement in Diagnosis, Prognosis and Therapy Monitoring

This review will focus on the elements of the skin’s immune system, immune cells and/or non-immune cells that support immune mechanisms, molecules with immune origin and/or immune functions that are involved in skin carcinogenesis. All these immune elements are compulsory in the development of skin tumors and/or sustainability of the neoplastic process. In this light, recent data gathered in this review will acknowledge all immune elements that contribute to skin tumorigenesis; moreover, they can serve as immune biomarkers. These immune markers can contribute to the diagnostic improvement, prognosis forecast, therapy monitoring, and even personalized therapeutical approach in skin cancer. Immune processes that sustain tumorigenesis in non-melanoma and melanoma skin cancers are described in the framework of recent data.

Biomarkers of metastatic melanoma

Melanoma, one of the most aggressive forms of human cancer, has undergone an alarming increase in incidence in recent years. Early detection is a prerequisite for proper diagnosis and therapy orientation. Soluble biomarkers are an important tool for early diagnosis. Markers that are associated with melanocyte functions imply the enzymes involved in melanin synthesis and the melanin-related metabolites. Proteins such as autocrine melanocyte cell growth factor and melanoma metastasis suppressor have gained attention in the biomarkers domain. The antimelanoma immune response elicited in patients can not only provide new biomarkers but important therapeutic approaches in specific treatments. All the molecules generated during the metastasis process, invasion of neighboring tissue, angiogenesis, invading lymphatic/blood vessels and establishing new tumors at a distant site, are targets for biomarker discovery.

Immune Parameters in The Prognosis and Therapy Monitoring of Cutaneous Melanoma Patients: Experience, Role, and Limitations

Cutaneous melanoma is an immune-dependent aggressive tumour. Up to our knowledge, there are no reports regarding immune parameters monitoring in longitudinal followup of melanoma patients. We report a followup for 36 months of the immune parameters of patients diagnosed in stages I–IV. The circulatory immune parameters comprised presurgery and postsurgery immune circulating peripheral cells and circulating intercommunicating cytokines. Based on our analysis, the prototype of the intratumor inflammatory infiltrate in a melanoma with good prognosis is composed of numerous T cells CD3+, few or even absent B cells CD20+, few or absent plasma cells CD138+, and present Langerhans cells CD1a+ or langerin+. Regarding circulatory immune cells, a marker that correlates with stage is CD4+/CD8+ ratio, and its decrease clearly indicates a worse prognosis of the disease. Moreover, even in advanced stages, patients that have an increased overall survival rate prove the increase of this ratio. The decrease in the circulating B lymphocytes with stage is balanced by an increase in circulating NK cells, a phenomenon observed in stage III. Out of all the tested cytokines in the followup, IL-6 level correlated with the patients survival, while in our study, IL-8, IL-10, and IL-12 did not correlate statistically in a significant way with overall survival, or relapse-free survival.

Microwave Synthesis, Basic Spectral and Biological Evaluation of Some Copper (II) Mesoporphyrinic Complexes

Cu(II) complexes with asymmetrical and symmetrical porphyrinic ligands were synthesized with superior yields using microwave irradiation. The paper presents the synthesis of 5-(3-hydroxyphenyl)-10,15,20-tris-(4-carboxymethylphenyl)-21,23-Cu(II)-porphine in comparison to its symmetrical complex 5,10,15,20-meso-tetrakis-(4-carboxy-methylphenyl)-21,23-Cu(II) porphine. The two compounds were characterized by FT-IR, UV–Vis and EPR spectroscopy, which fully confirmed the structures. The spectral molecular absorption properties of the porphyrinic complexes were studied in organic solvents (methanol, ethanol, iso-propanol, dimethyl sulfoxide, dimethylformamide and methylene chloride), and the influence of the solvent polarity on the absorbance maxima is described. In order to establish their future potential in biomedical applications preliminary toxicological studies consisting of viability and proliferation of standard tumor cell lines (MCF7 and B16) testing was performed. The obtained results indicate a low toxicity for both compounds and further recommends them for testing in light activation protocols.

Chemokines in the Melanoma Metastasis Biomarkers Portrait

Skin tumorigenesis is linked to inflammatory chemokines accumulation that can induce cancer-associated immune-suppression. Deregulation of the CXCR4/CXCL12 axis was reported in melanoma tumorigenesis while also linked to BRAF mutation. Some chemokine-receptor patterns can direct the organ-specific metastasis. CXCL10 can help to prognosticate high-risk patients as it is a chemokine that differentiated patients with vs. metastasis free ones. Besides serum/plasma, chemokine identification in the cerebrospinal fluid of melanoma patients can indicate brain metastasis. Interplay between suppressed and elevated chemokines in cerebrospinal fluid can pinpoint an aggressive melanoma brain metastasis. Chemokines are gaining rapid momentum in the biomarker discovery domain aiding melanoma prognosis and high-risk patients’ stratification.

Synthetic porphyrins in experimental photodynamic therapy induce a different antitumoral effect

The aim of the study was to investigate the influence of discrete structural differences in synthetic porphyrins on the antineoplastic effect induced in myelocitic cell line K562 after experimental photodynamic therapy procedure. For this study, we used 5,10,15,20-tetra(1-naphthyl)porphyrin, 5,10,15,20-tetra(4-sulfonatophenyl)porphyrin and their Zn complexes. For all these compounds, the following photodynamic therapy parameters were optimized: cell concentration, dye loading concentration, loading time and irradiation procedure. The non-toxic doses of porphyrins were different according to their structure: for 5,10,15,20-tetra(1-naphthyl)porphyrin compounds, the dose was of 10 μg.mL-1, and for 5,10,15,20-tetra(4-sulfonatophenyl)porphyrin compounds, the dose was 20 μg.mL-1. Cell functionality was assessed as membrane integrity, viability, and number of metabolically active cells. Photodynamic cell degradation kinetics showed that during irradiation tumor cells are actively destroyed, in contrast to unloaded cells. K562 cells loaded with the above-mentioned compounds and subjected to irradiation, displayed lower proliferative capacity compared to the control, for a period of 72 h after irradiation. The proliferative capacity of K562 cell line was more strongly inhibited by the 5,10,15,20-tetra(1-naphthyl)porphyrin family of compounds than by the 5,10,15,20-tetra(4-sulfonatophenyl)porphyrin family. The tested synthetic porphyrins showed effective antineoplastic activity against K562 leukemia cells in our in vitro photodynamic therapy experimental model.

Variations in the expression of TIMP1, TIMP2 and TIMP3 in cutaneous melanoma with regression and their possible function as prognostic predictors

Regression in melanoma is a frequent biological event of uncertain prognostic value as the lesion exhibits heterogeneous phenotypical features, both at the morphological and immunohistochemical level. In the present study, we examined the expression of tissue inhibitors of metalloproteinases (TIMP1, TIMP2 and TIMP3) in melanoma with regression. We specifically examined the expression levels of these TIMPs in regressed components (RC) and non-regressed components (NRC) of the tumor and compared their expression levels with those in non-regressed melanomas. We found that TIMP1 was overexpressed in the NRC of melanomas with partial regression (PR) compared with the NRC in melanomas with segmental regression (SR) (P=0.011). TIMP2 was overexpressed in the NRC of melanomas with PR compared with the NRC in melanomas with SR (PR/SR, P=0.009); or compared with the NRC in melanomas with simultaneous SR-PR (P=0.002); or compared with melanomas without regression (absence of regression) (P=0.037). Moreover, TIMP3 was overexpressed in the NRC of all melanomas with SR as compared to the RC component (P=0.007). Our findings on the differential expression of TIMP1, TIMP2 and TIMP3 in melanomas with regression support the hypothesis that the morphological differences identified in the melanoma regression spectrum may have a correlation with prognosis. This may explain the controversial findings within the literature concerning the biological and prognostic role of regression in melanoma.