Adeno-associated virus vector hydrogel formulations for brain cancer gene therapy applications.

Gelatin-based formulations are utilized in neurosurgical procedures, with Medisponge® serving as an illustration of a secure and biocompatible hemostatic formulation. Noteworthy are combined hemostatic products that integrate pharmacological agents with gelatin. Gelatin matrices, which host biologically active substances, provide a platform for a variety of molecules. Biopolymers function as carriers for chemicals and genes, a facet particularly pertinent in brain cancer therapy, as gene therapy complement conventional approaches. The registration of Zolgensma underscores the efficacy of rAAV vectors in therapeutic gene delivery to the CNS. rAAVs, renowned for their safety, stability, and neuron-targeting capabilities, predominate in CNS gene therapy studies. The effectiveness of rAAV vector therapy varies based on the serotype and administration route. Local gene therapy employing hydrogel (e.g., post-tumor resection) enables the circumvention of the blood-brain barrier and restricts formulation diffusion. This study formulates gelatin rAAV gene formulations and evaluates vector transduction potential. Transduction efficiency was assessed using ex vivo mouse brains and in vitro cancer cell lines. In vitro, the transduction of rAAV vectors in gelatin matrices was quantified through qPCR, measuring the itr and Gfp expression. rAAVDJ and rAAV2 demonstrated superior transduction in ex vivo and in vitro models. Among the cell lines tested (Hs683, B16-F10, NIH:OVCAR-3), gelatin matrix F1 exhibited selective transduction, particularly with Hs683 human glioma cells, surpassing the performance Medisponge®. This research highlights the exploration of local brain cancer therapy, emphasizing the potential of gelatin as an rAAV vector carrier for gene therapy. The functional transduction activity of gelatin rAAV formulations is demonstrated.

Profiling of microRNA as a tool to introduce rAAV vectors in gene therapy of breast cancer: A preliminary report.

BACKGROUND: Despite the wide range of diagnostic and therapeutic methods, breast cancer is responsible for many deaths each year. One of the original and novel cancer therapeutic approaches is gene therapy based on recombinant adeno-associated viral vectors. Among the molecular factors with the potential to become useful diagnostic biomarkers, microRNA (miRNA) molecules are being considered for personalized therapies. OBJECTIVES: The aim of the study was to examine the utility of miRNA profiling in the design of personalized recombinant adeno-associated virus (rAAV)-based gene therapy for breast cancer patients. MATERIAL AND METHODS: The analysis of 754 miRNAs in 7 breast cancer samples and control samples was performed using real-time polymerase chain reaction (PCR) based on TaqMan® Low-density Array (TLDA) cards. Online repositories were used to explore the relationship between miRNAs and genes encoding rAAV receptors (KIAA0319L, HSPG2, FGFR1, c-MET, PDGFRA, ITGB5, and RPSA). Then, we performed a comparative analysis of the results to examine the possibility of using miRNA profiling in the design of rAAV-based therapeutic protocols. RESULTS: Fifty-two percent of tested miRNAs were noted in at least 1 analyzed breast cancer and control tissue. Thirteen miRNAs were selected due to being outliers in the tested samples. In total, 155 miRNAs targeted genes encoding rAAV receptors in the tested samples (29 miRNAs for KIAA0319L, 60 miRNAs for c-MET, 31 miRNAs for HSPG2, 43 miRNAs for FGFR1, 36 miRNAs for PDGFRA, 18 miRNAs for RPSA, and 25 miRNAs for ITGB5). The expression of the selected miRNAs was not homogeneous across the 7 samples. CONCLUSION: Profiling of microRNA could be a significant factor in the design of rAAV-based personalized gene therapy for breast cancer patients.

The Activity of Vitamin C Against Ovarian Cancer Cells Is Enhanced by Hyperthermia.

BACKGROUND/AIM: Vitamin C is essential for the proper functioning of the human body and plays a crucial role in many biological processes as a cofactor for enzymes. The anticancer activity of vitamin C has been indicated for years. Hyperthermia used in clinics allows increasing the effectiveness of anticancer therapies and may also be useful in enhancing the action of other substances. The purpose of this study was to enhance the anticancer activity of vitamin C through hyperthermia against ovarian cancer cells. MATERIALS AND METHODS: The ovarian cancer cell lines Caov-3, NIH:OVCAR-3, and human fibroblasts CCD-1064Sk were tested in the present study. Vitamin C was used in the following concentrations: 0.24, 2.50 and 5.25 mM. Each of the selected concentrations was combined with the different temperatures (37°C, 40°C and 43°C). Cell survival, adhesion and changes at the molecular level were assessed. RESULTS: The obtained results revealed that hyperthermia enhances the anticancer activity of vitamin C. Ovarian cancer cells showed greater sensitivity to vitamin C at elevated temperatures. Cells may have different sensitivity to vitamin C due to the activation of different gene signatures associated with redox reactions and apoptosis, therefore we examined the following genes: BCAP31, BCL2L13, BID, CASP7, FADD and HTRA2. The increase in expression of these genes in cancer cells generated a stronger proapoptotic response. CONCLUSION: The present study showed that hyperthermia enhanced the anticancer activity of vitamin C in vitro.

The angiogenic activity of ascites in the course of ovarian cancer as a marker of disease progression.

Ovarian cancer cells are able to create invasive implants in the peritoneum and their growth is directly associated with the angiogenetic potential. This effect is probably stimulated by vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8), which are both found in ascites. The aim of this study was to assess the influence of ascites produced by ovarian cancer on the angiogenesis. Peritoneal fluid was collected from patients with advanced ovarian cancer; cancer cells were separated from CD45+ leukocytes. Angiogenesis was assessed in mice, after intradermal injection of full cellular suspension together with supernatant or phosphate buffered saline, purified cancer cells suspension, or CD45+ leukocytes suspension. The angiogenesis index (AI) was assessed after 72 hours. VEGF and Il-8 were measured in the supernatant and cellular suspension. AI was the highest in the isolated cancer cells suspensions as well in the group stimulated with supernatant. Both VEGF and IL-8 were high in supernatants from ascites rich in cancer cells (>45%). A significant correlation was revealed between IL-8 concentration and AI. We conclude that ascites in patients with advanced ovarian cancer stimulates angiogenesis and this mechanism is dependent mostly on cancer cells activity and enhanced by cooperation with infiltrating leukocytes.

Supplementation with hydrocortisone on the 3rd-5th day following dexamethasone premedicated chemotherapy eliminated severe dizziness and postural hypotension.

We present the case of a 60 year-old woman with a stage III fallopian tube cancer submitted to hysterectomy and bilateral salpingo-oophorectomy with partial omenectomy, followed by six courses of chemotherapy and revision surgery. After each course of chemotherapy (paclitaxel + carboplatin) preceded by premedication with dexamethasone, she suffered from side- -effects, of which the most unpleasant was severe dizziness appearing on the third, fourth and fifth day following the listed combination of drugs. It was revealed that dizziness with concomitant sweating and rapid pulse, noted in the standing position, was combined with marked postural hypotension. Considering the possibility of a temporary pituitary-adrenal axis suppression caused by premedication with a very large dose of dexamethasone, during those three days she was supplemented with small doses of hydrocortisone, which caused almost complete disappearance of the mentioned symptoms. Our conclusion is that postural hypotension causing severe dizziness initially linked with chemotherapeutic drugs can be eliminated or markedly reduced by three days supplementation with hydrocortisone applied after the expected wash out of the dexamethasone.

[Malignant ovarian germ cell tumors--clinical characteristics and analysis of outcomes]. / Zlosliwe guzy germinalne jajnika--charakterystyka grupy chorych i analiza wyników leczenia.

OBJECTIVES: Presentation of a group of patients with diagnosed malignant ovarian germ cell tumors (MOGCT), determination of prognostic factors and outcome analysis. MATERIAL AND METHODS: We selected patients with diagnosed malignant ovarian germ cell tumors from the patient registry of Cancer Center in Warsaw from 1990 to 2001. We analyzed clinical and pathological features of the study group, as well as methods and results of treatment. RESULTS: We collected documentation of 83 patients. Most were diagnosed with dysgerminoma and immature teratoma in the early stages of development. 73 patients received adjuvant chemotherapy after surgery At the end of the first line of treatment complete response was achieved in 63 patients (75.9%). Time to recurrence ranged from 25 to 518 days (mean 176 days). The most common site of recurrence was the true pelvis. The five-year overall survival was 62.7%. Significant favorable prognostic factor was early stage of disease and the histological diagnosis of dysgerminoma. From the 46 women after fertility-sparing surgery, 8 became pregnant. CONCLUSIONS: MOGCT are a group of potentially curable, yet very aggressive malignant ovarian tumors. The main condition for obtaining good results is quick diagnosis and appropriate treatment, usually surgery associated with multidrug chemotherapy The stage of the disease remains the most important prognostic factor. Patients diagnosed with dysgerminoma are a separate group with very good prognosis.

Clinical significance of serum CA-125 and soluble tumor necrosis factor receptor type I in cervical adenocarcinoma patients.

HYPOTHESIS: The purpose of this study was to answer the question whether the measurement of the pretreatment tumor markers and cytokine levels would be of clinical use in patients with cervical adenocarcinoma. METHODS: CA-125, carcinoembryonic antigen (CEA), and squamous cell carcinoma (SCC), as well as interleukin 6 (IL-6), IL-8, vascular endothelial growth factor, IL-1 receptor antagonist, soluble tumor necrosis factor receptor type I (sTNF RI), and sTNF RII, were assessed in the sera of 120 cervical adenocarcinoma patients. RESULTS: CA-125 presented a better diagnostic sensitivity than did CEA and SCC, whereas the concentration of most cytokines, except for sTNF RII, revealed higher sensitivity, than did the standard tumor markers. The highest sensitivity was found for sTNF RI. The concentrations of the examined parameters were found to be significantly higher in patients with advanced stage (IIB-IV) as compared with patients with I-IIA stage. [Float1]Serum concentration of IL-6 was the only one that differs significantly, depending on the histological grade. During the 3-year follow-up, 25 patients relapsed, and 73 patients were disease-free. Significantly higher pretreatment serum concentrations of the examined parameters (except for SCC and IL-1 receptor antagonist) were found in patients who developed recurrences. Soluble tumor necrosis factor receptor type I and CA-125 were found to present the highest sensitivity, with areas under the receiver operating characteristic curve of 0.833 and 0.809, respectively. As the result of univariate analysis, CA-125, CEA, sTNF RII, IL-6, sTNF RI, and clinical stage were considered factors of poor prognosis. Multivariate analysis has proven that CA-125 and clinical stage were the only significant independent prognostic factors of the disease-free survival. CONCLUSION: CA-125 is an independent prognostic factor for disease-free survival. Our results have also demonstrated that sTNF RI is probably the most useful marker in cervical adenocarcinoma patients, especially in the early stages of disease.

Increased cancer risk of heterozygotes with NBS1 germline mutations in Poland.

It has been suggested based on familial data that Nijmegen breakage syndrome (NBS) heterozygotes have an increased risk of malignant tumors. We found 15 carriers of the 657del5 mutation and 8 carriers of the R215W molecular variant of the NBS1 gene among 1,289 consecutive patients from Central Poland with various cancers and only 10 and 4 such carriers, respectively, in 1,620 controls from this region. Most of the 657del5 mutation carriers were found among patients with melanoma (4/105), non-Hodgkin lymphoma (2/42) and breast cancer (4/224) and of the 234 patients with colorectal carcinoma 3 carried the 657del5 mutation and 3 others the R215W molecular variant. The frequencies of 657del5 mutation carriers among patients with melanoma and non-Hodgkin lymphoma and of R215W carriers in patients with colorectal cancer were significantly higher than in controls (p < 0.01, < 0.05 and < 0.05 respectively). The pooled frequencies of 657del5 and R215W mutations in all cancer patients were also significantly higher than in controls (p < 0.05). Two carriers of the 657del5 mutation had second primary tumors. Malignant tumors among parents and siblings of 657del5 mutation carriers (14/77) were twice more frequent than in population controls. Three carriers of this mutation (2 probands with melanoma) reported melanoma in relatives. These results suggest strongly that NBS1 heterozygosity may be associated with elevated risk of some cancers. Larger studies are needed to evaluate the impact of the high frequency of germline NBS1 mutations on the cancer burden in the Slav populations.

[Surgical treatment of an isolated liver metastasis from endometrial cancer]. / Leczony chirurgicznie przerzut raka endometrium do watroby.

One of the principal features of the malignant process is the ability of the tumor to form metastases. In case of endometrial cancer pelvic and paraaortic lymph nodes and ovaries are the most frequent sites of metastases. In some cases metastases to the distant sites like the lungs and liver occur. The diagnosis of stage IVB disease bears a poor prognosis. Only a form of palliative treatment is usually offered to those unfortunate woman. The authors present a case of successful surgical treatment an isolated liver metastasis from endometrial cancer.