Andrey Turchinovich - Academia.edu (original) (raw)

Papers by Andrey Turchinovich

Supplementary Table S1: Chi-Square tests to assess difference in distribution between CTC-positiv... more Supplementary Table S1: Chi-Square tests to assess difference in distribution between CTC-positive and CTC-negative patients with respect to clinico-pathological features. # Wilcoxon rank sum test, *logrank test, a data corresponding to the primary tumour, b median could not be calculated as number

XLS file, 28K, Distribution of clinical characteristics of MBC cases.

XLS file, 19K, Interaction of miRNA levels and age in cases-control comparison.

PDF file, 90K, Partial correlations between miRNAs.

XLS file, 15K, Inter-run control variations.

Purpose: The use of circulating tumor cells (CTC) as a prognostic marker in metastatic breast can... more Purpose: The use of circulating tumor cells (CTC) as a prognostic marker in metastatic breast cancer (MBC) has been well established. However, their efficacy and accuracy are still under scrutiny mainly because of methods of their enrichment and identification. We hypothesized that circulating miRNAs can predict the CTC status of patients with MBC, and tested for the same. Furthermore, we aimed at establishing a panel of circulating miRNAs capable of differentiating MBC cases from healthy controls.Experimental Design: Circulating miRNAs from plasma of CTC-positive and CTC-negative patients with MBC, and healthy controls, were profiled by TaqMan Human MicroRNA arrays. Candidates from the initial screen were validated in an extended cohort of 269 individuals (61 CTC-positive, 72 CTC-negative, 60 CTC-low MBC cases, and 76 controls).Results: CTC-positive had significantly higher levels of miR-141, miR-200a, miR-200b, miR-200c, miR-203, miR-210, miR-375, and miR-801 than CTC-negative MBC...

Centrosome amplification is a hallmark of virtually all types of cancers, including solid tumors ... more Centrosome amplification is a hallmark of virtually all types of cancers, including solid tumors and hematologic malignancies. Cancer cells with extra centrosomes use centrosome clustering (CC) to allow for successful division. Because normal cells do not rely on this mechanism, CC is regarded as a promising target to selectively eradicate cells harboring supernumerary centrosomes. To identify novel inhibitors of CC, we developed a cell-based high-throughput screen that reports differential drug cytotoxicity for isogenic cell populations with different centrosome contents. We identified CP-673451 and crenolanib, two chemically related compounds originally developed for the inhibition of platelet-derived growth factor receptor β (PDGFR-β), as robust inhibitors of CC with selective cytotoxicity for cells with extra centrosomes. We demonstrate that these compounds induce mitotic spindle multipolarity by activation of the actin-severing protein cofilin, leading to destabilization of the...

Representative fluorescence time-lapse microscopy comparing mitotic events of H2B-mCherry-ÃŽÂ±-tu... more Representative fluorescence time-lapse microscopy comparing mitotic events of H2B-mCherry-ÃŽÂ±-tubulin-EGFP- HeLa cells treated with 2 Ã,ÂµM CP-673451.

Screen result. For each library and concentration, compounds are divided into two groups, accordi... more Screen result. For each library and concentration, compounds are divided into two groups, according to readout significance (black, P < 0.05; grey P > 0.05) and sorted from highest to lowest CCI index. Compounds highlighted in bold represent a CCI index higher than 0.3, indicating that viability of cells with CA was significantly affected in comparison to cells with normal centrosome content (by approx. 20%). Relative overall viability of each drug-treated cell population in comparison to the corresponding DMSO-treated population (maximum viability = 1) is indicated by rel.Control or rel.Tet. Compounds that eradicated both cell populations, giving a false positive result are highlighted in red (i.e. Crizotinib).

Extended experimental procedures Supplementary Fig. S1. Effect of CP-673451 and Crenolanib on cen... more Extended experimental procedures Supplementary Fig. S1. Effect of CP-673451 and Crenolanib on centrosome amplification in U2OS cells. Supplementary Fig. S2. Crenolanib prolongs of mitotic duration. Supplementary Fig. S3. CP-673451 activates cofilin in a dose-dependent manner in a variety of cancer cell lines. Supplementary Fig. S4. Inhibition of cofilin phosphorylation by LIMKi3 and Damnacanthal. Supplementary Fig. S5. Immunoblot analysis showing transient overexpression of different cofilin constructs and representative images of transfection efficiencies in U2OS-EGFP-PLK4 cells. Supplementary Fig. S6. Validation of SSH knock-down by real-time PCR. Supplementary Fig. S7. Effect of CP-673451 on SSH1 phosphatase activity. Supplementary Fig. S8. Immunoblot analysis of different cell lines showing that CP-673451 increases Akt and MEK1/2 phosphorylation under normal cultivation conditions. Supplementary Table S2. Average percentages of multipolar telophases in various cell lines after 2...

International Journal of Molecular Sciences, 2020

One of the main disadvantages of using DNA microarrays for miRNA expression profiling is the inab... more One of the main disadvantages of using DNA microarrays for miRNA expression profiling is the inability of adequate comparison of expression values across different miRNAs. This leads to a large amount of miRNAs with high scores which are actually not expressed in examined samples, i.e., false positives. We propose a post-processing algorithm which performs scoring of miRNAs in the results of microarray analysis based on expression values, time of discovery of miRNA, and correlation level between the expressions of miRNA and corresponding pre-miRNA in considered samples. The algorithm was successfully validated by the comparison of the results of its application to miRNA microarray breast tumor samples with publicly available miRNA-seq breast tumor data. Additionally, we obtained possible reasons why miRNA can appear as a false positive in microarray study using paired miRNA sequencing and array data. The use of DNA microarrays for estimating miRNA expression profile is limited by se...

Scientific Reports, 2016

Despite a high sequence homology among four human RNAi-effectors Argonaute proteins and their cod... more Despite a high sequence homology among four human RNAi-effectors Argonaute proteins and their coding sequences, the efficiency of ectopic overexpression of AGO3 and AGO4 coding sequences in human cells is greatly reduced as compared to AGO1 and AGO2. While investigating this phenomenon, we documented the existence of previously uncharacterized mechanism of gene expression regulation, which is manifested in greatly varying basal transcription levels from the RNApolII promoters depending on the promoter-proximal downstream sequences. Specifically, we show that distinct overexpression of Argonaute coding sequences cannot be explained by mRNA degradation in the cytoplasm or nucleus, and exhibits on transcriptional level. Furthermore, the first 1000–2000 nt located immediately downstream the promoter had the most critical influence on ectopic gene overexpression. The transcription inhibiting effect, associated with those downstream sequences, subsided with increasing distance to the prom...

Progress in Histochemistry and Cytochemistry, 2016

MicroRNA (miRNA) is a class of small non-coding RNAs which mediate post-transcriptional gene sile... more MicroRNA (miRNA) is a class of small non-coding RNAs which mediate post-transcriptional gene silencing (PTGS) by sequence-specific inhibition of target mRNAs translation and/or lowering their half-lives in the cytoplasm. Together with their binding partners, Argonaute (AGO) proteins, miRNAs form cores of RNA-induced silencing complexes (RISC). Despite a substantial progress in understanding RISC structure, until recently little was known about its localization in the cell. This review is aimed to provide an overview of the emerging picture of miRNA and RISC localization and function both in the intracellular space and outside of the cell. In contrast to the common assumption that PTGS occurs in the cytoplasm, it was found to operate mainly on the membranes of the endoplasmic reticulum (ER). Besides ER membranes miRNAs were found in all main cellular compartments including nucleus, nucleolus and mitochondria where they regulate various processes including transcription, translation, alternative splicing and DNA repair. Moreover, a certain pool of miRNAs may not be associated with RISC and carry completely different functions. Finally, the discovery of cell-free miRNAs in all biological fluids suggests that miRNAs might also act as signaling molecules outside the cell, and may be utilized as biomarkers for a variety of diseases. In this review we discuss miRNA secretion mechanisms and possible pathways of cell-cell communication via miRNA-containing exosomes in vivo.

Frontiers in Genetics, 2013

Nuclease resistant extracellular miRNAs have been found in all known biological fluids. The biolo... more Nuclease resistant extracellular miRNAs have been found in all known biological fluids. The biological function of extracellular miRNAs remains questionable; however, strong evidence suggests that these miRNAs can be more than just byproducts of cellular activity. Some extracellular miRNA species might carry cell-cell signaling function during various physiological and pathological processes. In this review, we discuss the state-of-the-art in the field of intercellular miRNA transport and highlight current theories regarding the origin and the biological function of extracellular miRNAs.

International Journal of Molecular Sciences

The extracellular circulating microRNA (miR)-200 regulates epithelial-mesenchymal transition and,... more The extracellular circulating microRNA (miR)-200 regulates epithelial-mesenchymal transition and, thus, plays an essential role in the metastatic cascade and has shown itself to be a promising prognostic and predictive biomarker in metastatic breast cancer (MBC). Expression levels of the plasma miR-200 family were analyzed in relationship to systemic treatment, circulating tumor cells (CTC) count, progression-free survival (PFS), and overall survival (OS). Expression of miR-200a, miR-200b, miR-200c, miR-141, and miR-429, and CTC status (CTC-positive ≥ 5 CTC/7.5 mL) was assessed in 47 patients at baseline (BL), after the first completed cycle of a new line of systemic therapy (1C), and upon the progression of disease (PD). MiR-200a, miR-200b, and miR-141 expression was reduced at 1C compared to BL. Upon PD, all miR-200s were upregulated compared to 1C. At all timepoints, the levels of miR-200s were elevated in CTC-positive versus CTC-negative patients. Further, heightened miR-200s ex...

iScience

The extracellular RNA communication consortium (ERCC) is an NIH-funded program aiming to promote ... more The extracellular RNA communication consortium (ERCC) is an NIH-funded program aiming to promote the development of new technologies, resources, and knowledge about exRNAs and their carriers. After Phase 1 (2013-2018), Phase 2 of the program (ERCC2, 2019-2023) aims to fill critical gaps in knowledge and technology to enable rigorous and reproducible methods for separation and characterization of both bulk populations of exRNA carriers and single EVs. ERCC2 investigators are also developing new bioinformatic pipelines to promote data integration through the exRNA atlas database. ERCC2 has established several Working Groups (Resource Sharing, Reagent Development, Data Analysis and Coordination, Technology Development, nomenclature, and Scientific Outreach) to promote collaboration between ERCC2 members and the broader scientific community. We expect that ERCC2's current and future achievements will significantly improve our understanding of exRNA biology and the development of accurate and efficient exRNA-based diagnostic, prognostic, and theranostic biomarker assays.