scholarly journals In Vitro Biophysical and Biological Characterization of Lipid Nanoparticles Co-Encapsulating Oncosuppressors miR-199b-5p and miR-204-5p as Potentiators of Target Therapy in Metastatic Melanoma

2020 ◽  
Vol 21 (6) ◽  
pp. 1930 ◽  
Author(s):  
Luigi Fattore ◽  
Virginia Campani ◽  
Ciro Francesco Ruggiero ◽  
Valentina Salvati ◽  
Domenico Liguoro ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Melanoma Cell ◽  
Biological Fluids ◽  
Target Therapy ◽  
Lipid Nanoparticles ◽  
Mapk Signaling ◽  
Oncogenic Driver

Uncontrolled MAPK signaling is the main oncogenic driver in metastatic melanomas bearing mutations in BRAF kinase. These tumors are currently treated with the combination of BRAF/MEK inhibitors (MAPKi), but this therapy is plagued by drug resistance. In this context we recently discovered that several microRNAs are involved in the development of drug resistance. In particular miR-204-5p and miR-199b-5p were found to function as antagonists of resistance because their enforced overexpression is able to inhibit melanoma cell growth in vitro either alone or in combination with MAPKi. However, the use of miRNAs in therapy is hampered by their rapid degradation in serum and biological fluids, as well as by the poor intracellular uptake. Here, we developed lipid nanoparticles (LNPs) encapsulating miR-204-5p, miR-199b-5p individually or in combination. We obtained LNPs with mean diameters < 200 nm and high miRNA encapsulation efficiency. These formulations were tested in vitro on several melanoma cell lines sensitive to MAPKi or rendered drug resistant. Our results show that LNPs encapsulating combinations of the two oncosuppressor miRNAs are highly efficient in impairing melanoma cell proliferation and viability, affect key signaling pathways involved in melanoma cell survival, and potentiate the efficacy of drugs inhibiting BRAF and MEK. These results warrant further assessment of the anti-tumor efficacy of oncosuppressor miRNAs encapsulating LNPs in in vivo tumor models.

scholarly journals FRET Imaging Approaches for in Vitro and in Vivo Characterization of Synthetic Lipid Nanoparticles

2014 ◽  
Vol 11 (9) ◽  
pp. 3133-3144 ◽  
Author(s):  
Julien Gravier ◽  
Lucie Sancey ◽  
Samuli Hirsjärvi ◽  
Emilie Rustique ◽  
Catherine Passirani ◽  
...  
Keyword(s):  
Lipid Nanoparticles ◽  
In Vivo Characterization

Identification and Characterization of Metabolites of Irinotecan in-vivo and in-vitro matrixes by HPLC/LC-MS

2016 ◽  
Vol 2 (3) ◽  
pp. 211-218
Author(s):  
Nidhi Srivastava ◽  
Vishal Dubey ◽  
Madhumita Sengar ◽  
Rastogi Sameer
Keyword(s):  
Method Development ◽  
Biological Fluids ◽  
Parent Compound ◽  
Enzymatic Reaction ◽  
Liver Microsomes ◽  
Metabolite Identification ◽  
Identification And Characterization

In the present study metabolite identification and characterization has done by using HPLC and LC-MS. During method development various mobile phases have tried for identification of metabolites. The matrixes selected for in- vivo study were urine because nearly all the metabolites of irinotecan were obtained in it. The extraction mixtures have selected to retain maximum amount of analyte with less effort. During experiment four extraction solvents were used in six different concentrations out of which TBME suit our method. In-vitro study done by Human Liver microsomes by using Phosphate buffer (pH 7.4) and NADPH as co-factors for initiation of enzymatic reaction. Irinotecan is a prodrug that is converted in the liver to an active metabolite, SN-38. It is eliminate in Bile and Faeces and thus its dose reduced in Hepatic Failure. Irinotecan act by inhibiting Topoisomerase-1.It is the enzyme which nicks, introduces negative supercoils and reseals the DNA strand. Conventionally, drug metabolite identification in the past has usually been based on the comparison of ultraviolet (UV) spectral data and high-performance liquid chromatography (HPLC) retention times of isolated ‘unknown’ metabolites with those of synthesised standards. Such a method of detecting and characterising drug metabolites is an uncertain, time-consuming and expensive process, as well as affording very limited structural information. Furthermore, Phase I metabolism of a drug candidate often results in only minor structural modification of the parent compound; these minor changes can make it particularly difficult to determine suitable chromatographic conditions to effect HPLC separation of metabolites. This study describes contemporary approach to identification and characterization of xenobiotic metabolites in complex biological fluids derived from drug metabolism studies.

scholarly journals Composition and Clinical Significance of Exosomes in Tuberculosis: A Systematic Literature Review

2021 ◽  
Vol 10 (1) ◽  
pp. 145
Author(s):  
Fantahun Biadglegne ◽  
Brigitte König ◽  
Arne C. Rodloff ◽  
Anca Dorhoi ◽  
Ulrich Sack
Keyword(s):  
Vaccine Development ◽  
Biological Fluids ◽  
Diagnostic Tools ◽  
Major Health ◽  
Fast Detection ◽  
Isolation And Characterization ◽  
Culture Supernatants

Tuberculosis (TB) remains a major health issue worldwide. In order to contain TB infections, improved vaccines as well as accurate and reliable diagnostic tools are desirable. Exosomes are employed for the diagnosis of various diseases. At present, research on exosomes in TB is still at the preliminary stage. Recent studies have described isolation and characterization of Mycobacterium tuberculosis (Mtb) derived exosomes in vivo and in vitro. Mtb-derived exosomes (Mtbexo) may be critical for TB pathogenesis by delivering mycobacterial-derived components to the recipient cells. Proteomic and transcriptomic analysis of Mtbexo have revealed a variety of proteins and miRNA, which are utilized by the TB bacteria for pathogenesis. Exosomes have been isolated in body fluids, are amenable for fast detection, and could contribute as diagnostic or prognostic biomarker to disease control. Extraction of exosomes from biological fluids is essential for the exosome research and requires careful standardization for TB. In this review, we summarized the different studies on Mtbexo molecules, including protein and miRNA and the methods used to detect exosomes in biological fluids and cell culture supernatants. Thus, the detection of Mtbexo molecules in biological fluids may have a potential to expedite the diagnosis of TB infection. Moreover, the analysis of Mtbexo may generate new aspects in vaccine development.

scholarly journals EWS-FLI1 regulates and cooperates with core regulatory circuitry in Ewing sarcoma

2020 ◽  
Vol 48 (20) ◽  
pp. 11434-11451
Author(s):  
Xianping Shi ◽  
Yueyuan Zheng ◽  
Liling Jiang ◽  
Bo Zhou ◽  
Wei Yang ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Ewing Sarcoma ◽  
Gene Mutations ◽  
Mapk Signaling ◽  
Genome Wide ◽  
Regulatory Loop ◽  
Sarcoma Cells

Abstract Core regulatory circuitry (CRC)-dependent transcriptional network is critical for developmental tumors in children and adolescents carrying few gene mutations. However, whether and how CRC contributes to transcription regulation in Ewing sarcoma is unknown. Here, we identify and functionally validate a CRC ‘trio’ constituted by three transcription factors (TFs): KLF15, TCF4 and NKX2-2, in Ewing sarcoma cells. Epigenomic analyses demonstrate that EWS-FLI1, the primary fusion driver for this cancer, directly establishes super-enhancers of each of these three TFs to activate their transcription. In turn, KLF15, TCF4 and NKX2-2 co-bind to their own and each other's super-enhancers and promoters, forming an inter-connected auto-regulatory loop. Functionally, CRC factors contribute significantly to cell proliferation of Ewing sarcoma both in vitro and in vivo. Mechanistically, CRC factors exhibit prominent capacity of co-regulating the epigenome in cooperation with EWS-FLI1, occupying 77.2% of promoters and 55.6% of enhancers genome-wide. Downstream, CRC TFs coordinately regulate gene expression networks in Ewing sarcoma, controlling important signaling pathways for cancer, such as lipid metabolism pathway, PI3K/AKT and MAPK signaling pathways. Together, molecular characterization of the oncogenic CRC model advances our understanding of the biology of Ewing sarcoma. Moreover, CRC-downstream genes and signaling pathways may contain potential therapeutic targets for this malignancy.

scholarly journals NCAM1 (CD56) promotes leukemogenesis and confers drug resistance in AML

Blood ◽  
2019 ◽  
Vol 133 (21) ◽  
pp. 2305-2319 ◽  
Author(s):  
Daniel Sasca ◽  
Jakub Szybinski ◽  
Andrea Schüler ◽  
Viral Shah ◽  
Jan Heidelberger ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Stress Resistance ◽  
Residual Disease ◽  
Mapk Pathway ◽  
Mapk Signaling ◽  
Aberrant Expression ◽  
Genotoxic Agents ◽  
The Impact

Abstract Neural cell adhesion molecule 1 (NCAM1; CD56) is expressed in up to 20% of acute myeloid leukemia (AML) patients. NCAM1 is widely used as a marker of minimal residual disease; however, the biological function of NCAM1 in AML remains elusive. In this study, we investigated the impact of NCAM1 expression on leukemogenesis, drug resistance, and its role as a biomarker to guide therapy. Beside t(8;21) leukemia, NCAM1 expression was found in most molecular AML subgroups at highly heterogeneous expression levels. Using complementary genetic strategies, we demonstrated an essential role of NCAM1 in the regulation of cell survival and stress resistance. Perturbation of NCAM1 induced cell death or differentiation and sensitized leukemic blasts toward genotoxic agents in vitro and in vivo. Furthermore, Ncam1 was highly expressed in leukemic progenitor cells in a murine leukemia model, and genetic depletion of Ncam1 prolonged disease latency and significantly reduced leukemia-initiating cells upon serial transplantation. To further analyze the mechanism of the NCAM1-associated phenotype, we performed phosphoproteomics and transcriptomics in different AML cell lines. NCAM1 expression strongly associated with constitutive activation of the MAPK-signaling pathway, regulation of apoptosis, or glycolysis. Pharmacological inhibition of MEK1/2 specifically inhibited proliferation and sensitized NCAM1+ AML cells to chemotherapy. In summary, our data demonstrate that aberrant expression of NCAM1 is involved in the maintenance of leukemic stem cells and confers stress resistance, likely due to activation of the MAPK pathway. Targeting MEK1/2 sensitizes AML blasts to genotoxic agents, indicating a role for NCAM1 as a biomarker to guide AML treatment.

scholarly journals LncRNA CCAT1 Enhances Chemoresistance in Hepatocellular Carcinoma by Targeting QKI-5

2020 ◽  
Author(s):  
Jing Shi ◽  
Cao Guo ◽  
Junli Ma
Keyword(s):  
Hepatocellular Carcinoma ◽  
Drug Resistance ◽  
Signaling Pathway ◽  
P38 Mapk ◽  
Tumor Model ◽  
Mapk Signaling ◽  
Mapk Signaling Pathway ◽  
Promising Alternative

Abstract Background: A major reason for treatment failure of cancer is acquisition of drug resistance. The specific mechanisms underlying hepatocellular carcinoma (HCC) chemoresistance need to be fully elucidated. lncRNAs involve in drug resistance in some cancers, however, the exact functions of lncRNA colon cancer-associated transcript 1 (CCAT1) in oxaliplatin resistance in HCC need to be elucidated.Methods: Functional analysis of CCAT1 on oxaliplatin sensitivity was performed in HCC cell lines HCCLM3 and HepG2, and in a subcutaneous tumor model receiving OXA treatment. Furthermore, the downstream signaling targets of CCAT1 in HCC were explored. Results: CCAT1 promoted HCC proliferation and reduced the apoptosis induced by oxaliplatin. Knockout of CCAT1 could increased chemosensitivity in vitro and in vivo. Further study found that QKI-5 was an important mediator and blocking of QKI-5/p38 MAPK signaling pathway enhanced oxaliplatin sensitivity.Conclusions: CCAT1 promoted proliferation and oxaliplatin resistance by QKI-5/p38 MAPK signaling pathway in HCC. Targeting CCAT1 in combination with chemotherapeutics may be a promising alternative to reverse drug resistance in HCC treatment.

Phytochemical-Mediated Glioma Targeted Treatment: Drug Resistance and Novel Delivery Systems

2020 ◽  
Vol 27 (4) ◽  
pp. 599-629 ◽  
Author(s):  
Hang Cao ◽  
Xuejun Li ◽  
Feiyifan Wang ◽  
Yueqi Zhang ◽  
Yi Xiong ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Side Effects ◽  
Target Therapy ◽  
Delivery Systems ◽  
Research Strategy ◽  
Satisfactory Outcome ◽  
Dismal Prognosis ◽  
Novel Drug

Glioma, especially its most malignant type, Glioblastoma (GBM), is the most common and the most aggressive malignant tumour in the central nervous system. Currently, we have no specific therapies that can significantly improve its dismal prognosis. Recent studies have reported promising in vitro experimental results of several novel glioma-targeting drugs; these studies are encouraging to both researchers and patients. However, clinical trials have revealed that novel compounds that focus on a single, clear glioma genetic alteration may not achieve a satisfactory outcome or have side effects that are unbearable. Based on this consensus, phytochemicals that exhibit multiple bioactivities have recently attracted much attention. Traditional Chinese medicine and traditional Indian medicine (Ayurveda) have shown that phytocompounds inhibit glioma angiogenesis, cancer stem cells and tumour proliferation; these results suggest a novel drug therapeutic strategy. However, single phytocompounds or their direct usage may not reverse comprehensive malignancy due to poor histological penetrability or relatively unsatisfactory in vivo efficiency. Recent research that has employed temozolomide combination treatment and Nanoparticles (NPs) with phytocompounds has revealed a powerful dual-target therapy and a high blood-brain barrier penetrability, which is accompanied by low side effects and strong specific targeting. This review is focused on major phytocompounds that have contributed to glioma-targeting treatment in recent years and their role in drug resistance inhibition, as well as novel drug delivery systems for clinical strategies. Lastly, we summarize a possible research strategy for the future.

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