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Aashir Awan, Ph.D.

Molecular & Cell Biologist

Personal Bio

During my training as a cell biologist, I have worked with two animal model systems (mouse and Tetrahymena) as well as having cell culturing experience, thereby becoming well-acquainted with a wide range of techniques and protocols (including but not limited to qPCR, cloning, antibody generation/purification, western blots, immunoassays, stem cell biology, primary cell culture, etc) in order to help answer key biological questions, ranging from basic cell biology (e.g. signal transduction) to finding and validating potential cancer targets.  I started out doing research in primary cilia where we did extensive characterizations of protein complexes (e.g. motor proteins transporting ciliary cargo) using an in-house generated antibody for immunoassays as well as using RNAi techniques.  And, during my first postdoctoral position, I characterized some of the signal transduction pathways present in human embryonic stem cells and in the pathology of ovarian cancer.  My second postdoc in Spain was trying to further delineate the hedgehog signal transduction pathway in different cell lines.  And, more recently, my postdoctoral work at Vanderbilt University centered on studying mouse CNS development and medulloblastoma tumorigenesis using techniques like immunofluorescence microscopy and by studying mouse knockouts.  As such, I became intimately involved in doing ex vivo experiments on primary cultures of cerebellar dissections and testing cancer drugs targeted to specific signal transduction proteins.

Aashir Awan, Ph.D.

Molecular & Cell Biologist

 

(203) 770 6402

http://www.linkedin.com/in/aashirawan

  

Techniques Summary

 

• Molecular biology (qPCR/gene cards, gel electrophoresis, lentivirus generation/infections, transfections, RNAi, etc.);
• Cell biology/biochemistry (culturing of primary human cell lines (including human embryonic stem cells), in vivo mouse modeling, ex vivo and in vitro-assays, western blots, antibody purification, elisas, etc.);
• Immunohistochemistry and immunofluorescence microscopy;

 

Education

 

Albert Einstein College of Medicine, Bronx, NY (2006)

PhD in Cell Biology, Advisor: Peter Satir

 

Albert Einstein College of Medicine, Bronx, NY (1999)

MS in Cell Biology, Advisor: Mitchell Bernstein

 

Johns Hopkins University, Baltimore, MD (1994)

B.A. Biology, Minor Hispanic Studies

 

Research Experience

 

07/15-07/16     Contract Research Scientist (In Vivo), Boehringer Ingelheim (Ridgefield, CT)

 

• Investigation of signal transduction pathways (e.g. hypoxia, GPCR) in pre-clinical/drug discovery phase of inflammatory bowel diseases (IBD) treatments using mouse modeling to determine the optimal time course/concentration of tested compounds;
• Conducting in vivo experiments testing the efficacy of inhibitors/drugs (e.g. small molecules against RORgt, CD226, etc.) or antibodies (e.g. p19/p40 against IL12/IL23) in key components of the immunology pathway using plasma/fecal/tissue samples from IBD mouse modeling experiments for gene expression (i.e. RNA isolation using automated liquid handling, cDNA synthesis, 384-well microfluidic gene cards), elisa as well as the subsequent data analysis (GraphPad software) of biomarkers;
• Analysis of cytokine levels of stimulations (e.g. CD3 antibody, curdlan, etc.) of mouse splenocyte cultures using ex vivo cultures (eg under hypoxia vs normoxia, acidic conditions, using small molecule inhibitors, etc.) for msd elisa and RNA analysis.

 

09/13-09/14     Associate Research Scientist, Yale School of Medicine (New Haven, CT)

 

• Development of approaches to targeted therapies for human salivary adenoid cystic carcinoma (ACC) using mammalian culturing, immunofluorescence (IF) microscopy, qPCR, mouse modeling, RNAi, SDS-PAGE western blots and bioinformatics analysis;
• Characterization of a cancer testis antigen, Ropn1b, as biomarker of salivary ACC/triple negative breast cancer cells and demonstrated a role for a Ropn1b peptide inhibitor’s ability to disrupt cell adhesion of cultured cancer line, HCC1599;
• Culturing the first primary human salivary ACC cell lines (ACCX6, ACC33) from patient-derived samples (using a modified stem cell media) and validated using biomarker analysis;
• Analyzed specific stem cell markers using siRNA (against Sox10, FABP7, Notch1) or small molecules (SBF160) in proliferation and qPCR assays of human salivary ACC cell lines.

 

04/11-04/12    Oncology & Development Postdoc, Vanderbilt University (Nashville, TN)

 

• Established the role of YAP1 in the homeostasis of the choroid plexus using immunohistochemistry and IF imaging;
• Showed YAP1’s role in medulloblastoma by cloning YAP GFP-constructs for retrovirus production and expression into cultured mouse cerebellar granule neuron precursors (CGNPs) for ex vivo assays (SDS-PAGE, western blots, IF microscopy) to evaluate proliferation and survival;
• Demonstrated a role for HIF1a during cerebellar development (under normoxic conditions) using ex vivo cultures (mouse CGNPs) with small molecule inhibitors for ex vivo assays;
• Validated the role of the p38 MAPK pathway in medulloblastoma tumorigenesis by evaluating apoptosis (i.e. caspase activity) using in vitro immunoassays of cultured human Daoy cell line using a small molecule inhibitor;
• Establishing primary mouse choroid plexus cultures using stem cell media.

 

03/09-03/11     Signal Transduction Fellow, Universidad Autonoma de Madrid (Madrid, Spain)

 

• Designed, subcloned and expressed 2 lentiviral constructs (recombinant protein expressions of GFP-tagged hedgehog receptors) into human HEK cells for infection of cultured CHO and U87 (human) glioblastoma cell lines to generate stable cell lines;
• Showed that the overexpression of hedgehog receptors resulted in massive filopodial outgrowths due to their parallel roles in the Cdc42 pathway using IF analysis.

 

09/06-01/09     Stem Cell & Ovarian Cancer Fellow, University of Copenhagen/Rigshospital (Copenhagen, Denmark)

 

• Validated cultured human embryonic stem cell lines (LRB 003, 010, 011) using a novel feeder-free stem cell media and characterized the hedgehog, wnt and PDGFa signal transduction systems in stem cell and blastocyst cilia;
• Used small molecules (e.g. smoothened agonist, SAG) for in vitro assays (immunofluorescence assays and qPCR) to show the functionality of hedgehog pathway in cultured human stem cells;
• Investigated the effects of small molecules (e.g. oxysterols) on the hedgehog and LXRa pathways using qPCR assays with mouse embryonic fibroblast (MEF) cell lines;
• Derived primary ovarian cell lines (from patient-derived samples) for comparison with human ovarian cancer cell lines (OVCAR3, SKOV3) using IF assays;
• Supervised 2 graduate students in the culturing conditions and subsequent molecular/ IF analysis of stem cells.

 

09/99-06/06     Cell Biology Graduate Research Assistant, Albert Einstein College of Medicine (Bronx, NY)

 

• Used PCR-based screening, southern/northern blotting, inverse PCR, cDNA/genomic libraries, and 5’-3’-RACE to subclone 5 different kinesin-like proteins in Tetrahymena for genomic sequencing;
• Generated/purified an antibody to Kin5 (utilizing peptide column chromatography) and demonstrated its ciliary localization using IF techniques;
• Devised and verified feasibility of an inducible RNAi system (including subcloning shRNA constructs into expression vectors) to knockdown genes in Tetrahymena;
• Demonstrated lethality of kin5 knockdown was due to loss of transport of membrane protein cargos in coimmunoprecipitation, SDS PAGE, western and motility/viability assays and establish role in Intraflagellar Transport;
• Worked on 2 protozoan species.

 

Honors

• 2015 NIH/NCI, 1R01 CA196200-01A1 (Investigator);
• 2009 Juan de la Cierva Fellowship (Spain);
• 2008 Danish Cancer Society Fellowship;
• 2006 Lundbeck Foundation Postdoctoral Fellowship (Denmark);
• 2005 Keith Porter Award (FASEB).

 

Publications

 

Awan A. (2016) Generating a successful Salivary Adenoid Cystic Carcinoma cell line. Challenges in Cancer Detection and Therapy (Accepted).

 

Vestergaard ML, Awan A, Warzecha CB, Christensen ST, Andersen CY (2014) Immunofluorescence and mRNA Analysis of Human Embryonic Stem Cells (hESCs) grown under feeder-free conditions. Methods Mol Biol 1307:123-140.

 

Egeberg DL, Lethan M, Manguso R, Schneider L, Awan A, Jørgensen TS, Byskov AG, Pedersen LB, Christensen ST (2012) Primary cilia and aberrant cell signaling in epithelial ovarian cancer. Cilia 1:1-15.

 

Guldal CG, Ahmad A, Korhunov A, Squatrito M, Awan A, Mainwaring LA, Bhatia B, Parathath SR, Nahle Z, Pfister S, Kenney AM (2012) An essential role for p38 MAPK downstream of Sonic hedgehog mitogenic signaling in cerebellar neural precursors. Acta Neuropathologica 123: 573-586.

 

Fernandez-L A, Squatrito M, Northcott P, Awan A, Holland EC, Taylor MD, Nahlé Z, Kenney AM (2011) Oncogenic YAP promotes radioresistance and genomic instability in medulloblastoma through IGF2-mediated Akt activation. Oncogene 379:1-15.

 

Awan A, Oliveri RO, Jensen PL, Christensen ST, Andersen CY (2010) Immunofluorescence and mRNA Analysis of Human Embryonic Stem Cells (hESCs) grown under feeder-free conditions. Methods Mol Biol. 584:195-210.

 

Awan A, Bell AJ, Satir P (2009) Kin5 knockdown in Tetrahymena thermophila using RNAi. PLoS One 4(3): e4873.

 

Veland IR, Awan A, Pedersen LB, Yoder BK, Christensen ST (2009) Primary cilia and signaling pathways in mammalian development, health and disease. Nephron Physiol. 111(3): 39-53.

 

Nielsen SK, Møllgård K, Clement CA, Veland IR, Awan A, Yoder BK, Novak I, Christensen ST (2008) Characterization of primary cilia and Hedgehog signalling during development of the human pancreas and in human pancreatic duct cancer cell lines. Dev Dyn 237(8): 2039-2052.

 

(Kiprilov¹ E, Awan¹ A), Desperat R, Velho M, Celment CA, Byskov AG, Andersen CY, Satir P, Bouhassira EE, Christensen ST, Hirsch RE (2008) Human embryonic stem cells in culture possess primary cilia with hedgehog signaling machinery. J Cell Biol. 180(5): 897-904.

(¹ Both Authors contributed equally)

 

Awan A, Bernstein M, Hamasaki T, Satir P (2004) Cloning and characterization of Kin5, a novel Tetrahymena ciliary kinesin II. Cell Motil Cytoskeleton 58(1):1-9.

 

Christensen ST, Guerra CF, Awan A, Wheatley DN, Satir P (2003) Insulin receptor-like proteins in Tetrahymena thermophila ciliary membranes. Curr Biol. 13(2):R50-R52.

 

Other Information

 

• US citizen;
• Reviewer for: Journal Cell Science, Journal Lipid Research;
• Member, American Association for Cancer Research;
• Volunteer, Tennessee Horticultural Society
• Native English speaker, fluent speaker/writer in Spanish, fluent speaker in Urdu.