Andrew Zlobin PhD
Molecular Biologist
Chicago,USA
Summary
Ph.D. with an extensive knowledge and expertise of Cellular & Molecular Biology and Biochemistry primarily in Breast Cancer and Immunology.
20 years of expertise in studying complex gene regulation using molecular-, genetic-, biochemical-, in vivo mouse model- based approaches. Next Generation Sequencing applications for transcriptome analysis including data analysis. Expertise evidenced by peer reviewed publications in the field of Oncology.
Overview
20
20
years of professional experience
Work History
Staff Scientist
University Pittsburgh
Pittsburgh, PA
06.2002 - Current
- Department of Medicine, Section of Rheumatology
- Investigation of Idiopathic inflammatory myopathies – dermatomyositis (DM) and inclusion body myositis (IBM), which were done by FACS sorting with expression analysis. Clinical DM and IBM samples (biopsy) were subjected to FACS, CD19+ cells and downstream CD27+IgD- and CD138+ cell subsets were used to identify major antibody secreting cell subsets. Expression of heavy and light chain and transcription factors were confirmed by RT-PCR. High-Throughput single-cell RNA sequencing analysis was performed for an in-depth characterization of this populations and determine its’ contribution to in situ humoral autoimmunity. Image analysis of multichannel immunofluorescent Confocal microscopy was used for the segmentation analysis of in vivo cellular interactions and to identify localization of specific immune cells subsets. NGS 10x platform used to characterize different cell populations and distribution of immune cell populations
- The same project as at Pitt University
Staff Scientist
University of Chicago
Chicago, IL
06.2020 - 06.2022
- Department of Medicine, Section of Rheumatology
- Investigation of idiopathic inflammatory myopathies – dermatomyositis (DM) and inclusion body myositis (IBM) were done by FACS sorting with expression analysis. Clinical DM and IBM samples (biopsy) were subjected to FACS, CD19+ cells and downstream CD27+IgD- and CD138+ cell subsets were used to identify major antibody secreting cell subsets. Expression of heavy and light chain and transcription factors were confirmed by RT-PCR. High-Throughput single-cell RNA sequencing analysis was performed for an in-depth characterization of this populations and determine its’ contribution to in situ humoral autoimmunity. Image analysis of multichannel immunofluorescent Confocal microscopy was used for characterization of the segmentation analysis of in vivo cellular interactions. NGS 10x platform used characterize different cell populations and distribution of immune cell populations
- The same project as at Pitt University
Senior Staff Scientist
Loyola University of Chicago
Maywood, IL
06.2018 - 06.2020
- Department of Physiology
- I’ve explored the mechanistic role and functional consequence of GSK-3b on the myofilament in the context of ischemia-induced dyssynchrony. Ischemia-induced dyssynchrony was modeled through left anterior descending coronary ligation resulting in myocardial infarction (MI) in WT and cardiomyocyte-specific GSK-5b KO mice. In WT mice, myofilament GSK5b decreased to KO levels 48-hours post-MI, corresponding to an increase in mechanical dyssynchrony. Strain analysis confirmed this dyssynchrony, and revealed mild dyssynchrony in KO mice at baseline, in which the posterior wall contracts prior to the anterior wall. We also found in WT mice higher levels of myofilament-GSK-5b in the anterior compared to posterior wall (which did not correlate with cytosolic GSK-5b). Mass spectrometry was used for this project.
Research Associate
Loyola University of Chicago
Maywood, IL
06.2009 - 06.2018
- Breast Cancer Division
- My two projects were focused on breast cancer. Projects were: “DAXX Suppresses Tumor-Initiating Cells in Estrogen Receptor-positive Breast Cancer following endocrine therapy” and “A Study of GSI inhibitor MK-0752 in Combination with Tamoxifen and Letrozole in Patients with Early Stage Breast Cancer Prior to Surgery.” In these clinical studies I focused on pathways and biomarkers modulation during treatment of patients. DAXX and Notch signaling pathways play an important role in tumor proliferation and progression. We hypothesized that short GSI MK-0752 treatment in combination with endocrine therapy will result in modulation of cancer cells proliferation. Analyzing FFPE specimens of patients with early stage ERα+ breast cancer we were able to indicated significant impact of GSI treatment on biomarkers. Expression of anti-proliferative and pro-apoptotic genes at the molecular level was significantly affected.
Research Associate
Loyola University of Chicago
Maywood, IL
06.2007 - 09.2009
- Skin Cancer
- Proapoptotic Noxa expression is regulated by E2F1 transcription factor in A375 - Human Metastatic Melanoma cell line after treatment with Apomine and Velcade. One of factors contributing to mechanism of apoptotic death in neoplastics treated with proteosome inhibitors is activation of Noxa, which is important in triggering apoptotic machinery. We demonstrate that activation of apoptosis caused by Apo or Velcade is dependent on E2F1 transcription factor binding to E-box of Noxa promoter. E2F1 Isi-RNA treatment of cells abrogate activation of Noxa. We used combination of methods like luciferase and ChIP assay, computer based prediction of binding site of Noxa promoter region, etc.
Research Associate
Loyola University of Chicago
Maywood, IL
06.2001 - 06.2007
- Department of Molecular Oncology
- Functions of histone modifications in regulation of Hox genes expression. The role of Intergenic RNA transcription and Cyclophylin 3s in the regulation of Hox gene expression. This project involves studying the functions of non-coding RNA transcripts and histone methylation in regulation of developmentally active abdA,B and Ubx genes. We’ve got a lot of very important results, based on those the model was proposed in which the Cyp3s functions as RNA sensors for the trithorax complex, intergenic RNA sequester Cyp3s that otherwise promote gene silencing through interaction with trithorax (TRX) and polycomb (PC) group regulatory complexes. Also we use ChIP assay to decipher involvement of histone lysine residues acetylation and methylation in regulation of Hox genes expression. Also RNA interference (siRNA approach) was used to evaluate regulation of abd A, B and UBX genes. Data are being published.
Research Associate
Loyola University of Chicago
Maywood, IL
07.1998 - 06.2001
- Department of Molecular Oncology
- Activation of apoptotic pathway in neoplastic cells using recombinant retrovirus vectors caring inducible apoptotic gene. I worked on the application of gene therapy approach to treatment of human neoplastic malignancies by activating apoptotic cell death. It has been shown that overexpression of Notch1 receptor protects transformed cell from apoptotic death. To downregulate the expression of the Notch1 gene and block the effects of its product I developed recombinant Retrovirus vectors carrying drug inducible antisense Notch1 cDNA. The Retroviruses were being used to transfect cancer cells and make them susceptible to drug treatment. Additionally, we were investigating mechanisms of action of Notch1 receptor and signal transduction pathway(s) upstream and downstream of this gene product. Use molecular biology techniques and flow cytometry for research.
Postdoctoral Research Associate
Virginia Commonwealth University
Richmond, VA
06.1996 - 06.1998
- Molecular Genetics Laboratory
- Molecular Genetics of Schizophrenia and ADHD, detecting the vulnerability locus for nicotine addition. The goals of research were to identify genes responsible for the vulnerability locus of schizophrenia, and Attention-Deficit Hyperactivity Disorder (ADHD). Detection of susceptibility genes for these psychiatric disorders was performed using a genome wide scan & a candidate gene approach, along with the utilization of linkage and association analyses.
Postdoctoral Research Associate
The Rockefeller University
New York, NY
06.1995 - 06.1996
- Department of the Biology of Addictive Diseases
- Investigating the role of dopamine transporter and receptors in developing cocaine addiction. This project involves studying the level of dopamine receptor and transporter mRNAs in different part of brain. I used molecular biology techniques like RT-PCR, RNase protection assay.
Education
Doctor of Philosophy - Molecular Virology
Institute of Virology, Academy of Science
Moscow, Russian Federation01-1981
Master’s of Science - Molecular Biology
Moscow State University
Moscow, Russian Federation05-1975
Skills
- Molecular work
- High Throughput Single Cell RNA Sequencing
- Chromatin-Immuno Precipitation assay (ChIP), Chip-SEQ analysis, Transcription co-factors, Histone methylation
- RNA interference: experience with developing, validating, and applying oligonucleotide and viral (si and sh) RNAi reagents, for transcript/protein knockdown
- Retro-Viral Vectors, transfection, On/Off system
- QPCR, PCR, library screening, cloning, transfection, transformations, making probes, Western/Southern/Northern hybridization, DNA sequencing
- Site-directed mutagenesis, gel mobility shift assay, RNase protection assay
- Flow cytometry (FACS) analysis and sorting
- Confocal microscopy
- Enzymology, protein purification: chromatography techniques, electrophoresis, enzyme assays, protein labeling, pulse chase
- Biochemical binding assays: ELISA, ligand blots, dot blots, immunoprecipitation, column, thin layer, gas chromatography
- Virology methods
- Expert in Mammalian cell culture, Spheroids cell culture, Pharmacologic drug treatment, Genetic manipulations to create stable and transient cell line development Virus mediated gene therapy, recombinant DNA techniques including transfection/transformation
- Profound computer literacy with PC, MAC, Linus OS, Proficient in Bioinformatics: NGS RNA-SEQ, Chip-SEQ (NGS) analysis (Galaxy platform, R-Studio, on-line software) NCBI (BLAST), EMBL, UCSC, Ensemble, Gene Card, SWISS-PROT, Uniprot, Motif finder, and Primer Design Statistics software –SPSS, Microsoft Office R platform
- Have supervisory, training and project management experience
- Animal work
- Experienced in managing and handling laboratory mouse experiments and maintenance of tumor line in vivo
- Trained in small animal surgery: peritoneal injections, oral gavage, subcutaneous/orthotopic xenograft transplantation, body weight and tumor volume measurements
- Performed animal tissue sectioning using cryostat and microtome for immunohistochemistry
- Isolated RNA and protein from frozen and paraffin embedded tissue sections
PUBLICATIONS
- 1. Ai L, Perez E, Asimes A, Kampanegstri T, Heroux M, Zlobin A, Hiske MA, Chung CS, Pak TR, Kirk JA. Binge Alcohol Exposure in Adolescence Impairs Normal Heart Growth. J Am Heart Assoc. 2020 May 5;9(9):e015611. doi: 10.1161/JAHA.119.015611. Epub 2020 Apr 22. PMID: 32319345
- 2. Reiter R, Wyatt D, Zlobin A, Piracha A, Ng J, Dingwall AK, Albain KS, Osipo C. DAXX Suppresses Tumor-Initiating Cells in Estrogen Receptor-Positive Breast Cancer Following Endocrine Therapy. Cancer Res. 2019 Oct 1;79(19):4965-4977. doi: 10.1158/0008-5472.CAN-19-0572. Epub 2019 Aug 6.
- 3. Pandya K, Wyatt D, Gallagher B, Bloodworth J, Shah D, Baker A, Zlobin A, Pannati A, Green AR, Ellis IO, Filipovic A, Sargent J, Rana A, Albain KS, Miele L, Denning M, Osipo C. PKCα Attenuates Jagged-1-Mediated Notch Signaling in ErbB-2-Positive Breast Cancer to Reverse Trastuzumab Resistance. Clin Cancer Res. 2016 Sep 8. pii: clincanres.0197.2015.
- 4. Baker AT, Zlobin A, Osipo C., Notch-EGFR/Her2 Bidirectional Crosstalk in Breast Cancer. Front Oncol. 2014 Dec 12;4:360. doi: 10.3389/fonc.2014.00360. eCollection 2014. Review.
- 5. Weber CE, Kothari AN, Wai PY, Li NY, Driver J, Zapf MA, Franzen CA, Gupta GN, Osipo C, Zlobin A, Syn WK, Zhang J, Kuo PC, Miz J. Osteopontin mediates an MZF1-TGF-β1-dependent transformation of mesenchymal cells into cancer-associated fibroblasts in breast cancer. Oncogene. 2014 Dec 22. doi: 10.1038/onc.2014.410.
- 6. Albain KS, Zlobin A, Covington K, et al. Identification of a Notch-driven Breast Cancer Stem Cell Gene Signature for Anti-Notch Therapy in an ER-Positive Presurgical Window Model. Oral presentation, SABCS 2014.
- 7. Olusakass-Kuprys K, Zlobin A, Osipo C, Gamma secretase inhibitors of Notch signaling. OncoTargets and Therapy, July 2013, 6:943-955,25
- 8. Zlobin A, Kuprys-Olusakass R, Shah D, Osipo C. Ratio of Notch receptors is critical for response to inhibition by a gamma-secretase inhibitor in triple negative breast cancer cells. Abstract, The 2013 San Antonio Breast Cancer Symposium.
- 9. KS. Albain, C Czerlains, A Zlobin, KR Covington, P Rajan, C Godellas, D Bova, SS Lo, P Robinson, S Sarker, ER Gaynor, R Cooper, G Aranha, K Czapliki B, Busby P, Rizzo T, Demuth P, Stiff S,AW Fuqua, and L Miele: Modulation of Cancer and Stem Cell Biomarkers by the Notch Inhibitor MK-0752 Added to Endocrine Therapy for Early Stage ER+ Breast Cancer. Cancer Research, February 9, 2012 71:1-5; doi:10.1158/0008-5472.SABC11-S1-5.
- 10. A Zlobin, R Olusakass-Kuprys, S Hodge M, O’Toole C, Ersahin, and C Osipo. Assessment of Notch Signaling Pathway Components as Biomarkers for Triple Negative Breast Cancer: Comparison of Triple Negative Breast Cancer Cell Lines and Human Breast Cancer Samples. Cancer Research, February 9, 2012 71:1-5; doi:10.1158/0008-5472.SABC11-S1-5.
- 11. C Osipo A, Baumgartner A, Zlobin, and L Miele. Notch-dependent Regulation of Novel Genes Associated with Trastuzumab Resistance. Cancer Research, December 17, 2012 72:P4-08-06; doi:10.1158/0008-5472.SABC12-P4-08-06.
- 12. Albain K, Czerlains, C, Rajan P, Zlobin A, Godellas C, Bova D, Lo SS, Robinson P, Sarker S, Gaynor ER, Cooper R,Aranha G, Czapliki K, Busby P, Rizzo P, Chissame M, Demuth T, Blackman S, Watters J, Stiff P, Fuqua SAW, Miele L: Combination of Notch Inhibitor MK-0752 and Endocrine Therapy for Early Satge ER+ Breast Cancer in a Presurgical Window Pilot Study. Cancer Res 2010, 70.
- 13. Tie F, Banerjee R, Stratton CA, Prasad-Sinha J, Stepanik V, Zlobin A, Diaz MO, Scacheri PC, Harte PJ. CBP-mediated acetylation of histone H3 lysine 27 antagonizes Drosophila Polycomb silencing. Development. 2009 Sep;136(15):3131-41.
- 14. Sonty L, Zlobin A, Ghoshal P, Zhang Q, Houde C, Weijzen S, Jiang Q, Nacheva E, Yagan D, Davis E, Gallego-Zouitina S, Catovsky D, Grogan T, Fisher RI, Miele L, Coignet LJ. Alteration of SMRT tumor suppressor function in transformed non-Hodgkin lymphomas. Cancer Res. 2005 Jun 1;65(11):4551-61.
- 15. Jang MS, Miao H, Carlesso N, Shelly L, Zlobin A, Darack N, Qin JZ, Nickoloff BJ, Miele L. Notch-1 regulates cell death independently of differentiation in murine erythroleukemia cells through multiple apoptosis and cell cycle pathways. J Cell Physiol. 2004 Jun; 199(3): 418-53.
- 16. Weijzen S, Zlobin A, Brad M, Miele L, Kast W. HPV16 E6 and E7 oncoproteins regulate Notch-1 expression and cooperate to induce transformation. J Cell Physiol. 2003 Mar; 194(3): 356-62.
- 17. Weijzen S, Rizzo P, Brad M, Vaishnav R, Zlobin A, Osborne B, Gottipati S, Aster J, Rudolf M, Hahn W, Kast W., Miele L. Activation of Notch-1 signaling maintains the neoplastic phenotype in human Ras-transformed cells. Nature of Medicine 2002 Sep;8(9): 979-86.
- 18. Cheng P, Zlobin A, Voglina V, Gottipatti S, Osborn B, Simel E, Miele L and Gabrilovich D. Notch-1 regulates NF-kB activity in hemopoietic progenitor cells. Journal of Immunology, 2001 Oct 15;167(8):1458-1467.
- 19. Zlobin A, Jang M, Miele L, Cozzo J. Toward the rational design of cell fate modifiers: Notch signaling as a target for novel biopharmaceuticals. Current Pharmaceutical Biotechnology, 2000, 1, 85-106.
- 20. Jang M, Zlobin A, Kast W, Miele L. (2000). Notch signaling as a target in multidomain cancer therapy. Current Opinion in Molecular Therapeutics 2000, Vol 2 No 1, pp 55-65.
- 21. Spangler R, Zhou Y, Maggos CE, Zlobin A, Ho A, Kreek MJ (1996). Dopamine antagonist and “binge” cocaine effects on rat opioid and dopamine transporter mRNAs. Neuroreport 1996 Sep 2; 7:13 2196-200.
PERSONAL
Russian, English – full professional proficiency, Russian language=Native proficiency, married, US citizen
Timeline
Staff Scientist
University of Chicago
06.2020 - 06.2022
Senior Staff Scientist
Loyola University of Chicago
06.2018 - 06.2020
Research Associate
Loyola University of Chicago
06.2009 - 06.2018
Research Associate
Loyola University of Chicago
06.2007 - 09.2009
Staff Scientist
University Pittsburgh
06.2002 - Current
Research Associate
Loyola University of Chicago
06.2001 - 06.2007
Research Associate
Loyola University of Chicago
07.1998 - 06.2001
Postdoctoral Research Associate
Virginia Commonwealth University
06.1996 - 06.1998
Postdoctoral Research Associate
The Rockefeller University
06.1995 - 06.1996
Master’s of Science - Molecular Biology
Moscow State University
Doctor of Philosophy - Molecular Virology
Institute of Virology, Academy of Science