Clinical Research Scientist

Mariana Figuera Losada, Ph. D.

*** ********* ***, ** *, Bronx, NY 10465

574-***-****, ********@*******.***

Protein biochemist and science writer interested in applying clinical research skills to contribute to the understanding of the molecular basis of diseases and the development of new drugs. Effective and adaptable team member with strong written and oral communication skills, problem solving and multi-tasking abilities.

RESEARCH EXPERIENCE

Montefiore Medical Center June, 2017-present

oClinical Research Coordinator Volunteer at the Sleep-Wake Disorders Center, Bronx, NY

oPerform the duties of a Clinical Research Coordinator for five different ongoing clinical trials taking place at the Sleep-Wake Disorders Center. These tasks include working with regulatory binders, creating source documents, assist with subject recruitment tasks, carry out pre-screening interviews and study visit procedures.

oOversight of all the tasks related to a sleep disorder prevalence study proposed by Dr. Imran Ahmed as a project for a student volunteering at the center. These tasks include: designing, writing and submitting the study protocol to the corresponding regulatory agencies, gathering the study data, analyzing the data and ensuring that all the study personnel have the appropriate training. Creation of cohorts of patients required for study using Clinical Looking Glass tool (Streamline Health-Montefiore Medical Center).

Albert Einstein College of Medicine, Bronx, NY April, 2015-June 2017

Associate at Dr. Vern Schramm laboratory, Department of Biochemistry

oElucidate the molecular and cellular mechanisms of the anticancer compound methylthio-DADMe-immucillin A (MT-DIA), a transition state analogue that inhibits 5’-methylthioadenosine phosphorylase (MTAP) with a Ki = 86 pM. MTAP is a key enzyme in the metabolism of 5’-methylthioadenosine (MTA) in humans and it plays a major role in polyamine metabolism and the salvage of adenine and methionine.

oStudy the mechanism of resistance to MT-DIA in a head and neck cancer cell line (FaDu cells).

TECHNICAL SKILLS:

Culture of multiple mammalian cancer cells lines including head and neck cancer cells, glioblastoma, osteosarcoma, colorectal carcinoma and triple negative breast cancer cells.

3D culture of cancer cells as spheroids and single cells embedded in Matrigel.

In vitro wound healing and matrix invasion assays.

Knockdown of protein expression in mammalian cells using shRNA and siRNA.

Lentivirus and retrovirus infection of mammalian cells.

Retrovirus packing, isolation and concentration. Viral titer determination using RT-PCR.

Pull-down assays to assess effects of MT-DIA on PRC2 complex composition.

Analysis of histone H3 and H4 methylation using Western-blotting.

Studied the effects of various compounds on DNA methylation in FaDu cells.

Flow cytometry for cell death analysis and cell sorting of GFP or RFP expressing cells.

Developed and carried out HPLC-based assays to identify enzymatic reaction products.

ORGANIZATIONAL SKILLS:

Setup a new fully functional and independent tissue culture room with biosafety level 2 certification.

Responsible for equipment maintenance and certification.

In charge of ordering reagents and supplies for the tissue culture room.

Johns Hopkins University, Baltimore, MD June, 2012-March, 2015

Research fellow at Dr. Barbara S. Slusher laboratory, Brain Science Institute

oResponsible for in vitro assays in a translational research project for HIV-associated neurocognitive disorders.

oDeveloped and implemented enzymatic, cell based and phenotypic assays to screen for potential inhibitors of neutral sphingomyelinase (nSMase), system xC-, glutaminase, positive allosteric modulators of metabotropic glutamate 1 receptor (mGluR1) and inhibitors of microglial cell activation.

TECHNICAL SKILLS:

Primary hippocampal and cortical neuronal cell culture. Primary and immortalized microglia and macrophage cell culture.

Tecan liquid handling robot Freedom EVO.

Intracellular calcium changes using a fluorescence imaging plate reader (FLIPR).

Fluorescence and radioactive (3H-labeled substrate) assays for nSMase and system xc-.

Measurement of glutamate release from microglia cells, macrophages and astrocytoma cells using a fluorescence coupled-reaction assay.

Protein overexpression in mammalian cells (HEK293).

MANAGEMENT AND ORGANIZATIONAL SKILLS:

In charge of one research technician and one undergraduate student.

Setup a full molecular biology/biochemistry laboratory from the ground up.

In charge of ordering reagents and supplies for my own project and the tissue culture room.

The Scripps Research Institute, Jupiter, FL September, 2008-April, 2012

Research Associate at Dr. Philip V. LoGrasso laboratory, Molecular Therapeutics Department

oCharacterized cJun N-terminal kinase (JNK) enzymatic activity and interaction with substrates.

oStudied role of JNK in cellular death induced by toxins, oxidative stress, endoplasmic reticulum stress and inflammatory cytokines providing an insight in the understanding of the roles of JNK in Parkinson’s disease and type II diabetes.

oStudied substrate specificity of JNK isoforms and splice variants.

oScreened libraries of small molecule compounds in high-throughput format.

TECHNICAL SKILLS:

Development and implementation of various assays based on absorbance, radioactive 33P-labeled substrate, homogeneous time-resolved fluorescence (HTRF), fluorescence resonance energy transfer (FRET), fluorescence polarization (FP) and cell based assays. These assays were used to screen for inhibitor compounds.

Improved expression and purification of JNK substrate, cJun by designing a fusion protein construct.

Biolayer interferometry (BLI) using the fortéBIO Octet Red instrument to study kinetics of interaction between JNK and various ligands.

Liquid handling instruments (pin tool and FRD).

Molecular biology and cell biology techniques: 2D electrophoresis, Western blotting, in-cell Westerns, ELISAs, immunoprecipitation, subcellular fractionation, in vitro and in vivo protein biotinylation, enzymatic digestion of proteins, cell staining for detection and quantitation of cell death, apoptosis, viability, reactive oxygen species and mitochondrial membrane potential dissipation.

MANAGEMENT AND ORGANIZATIONAL SKILLS:

Trained laboratory personnel and undergraduate students in the proper use of equipment and the execution of various experimental techniques.

Single-handedly packed, unpacked and moved the entire laboratory to a new location.

Setup a tissue culture room (biosafety level 2, BSL2) from the ground up.

In charge of maintenance, scheduling service and certification, and keeping track of service contracts for all the instruments in the laboratory, including an fortéBIO Octet red and a fast protein liquid chromatography AKTA FPLC (GE Healthcare Life Sciences).

In charge of ordering supplies and reagents using Sciquest system.

University of Notre Dame, Notre Dame, IN August, 2001-July, 2008

Graduate Student Researcher at Dr. F. J. Castellino laboratory, Department of Chemistry and Biochemistry

oCharacterized the effects of the interaction of streptococcal PAM protein-derived peptide VEK-30 with plasminogen (Pg).

oDiscovered residues in Pg domain kringle 2, key for VEK-30 binding.

oAscertained the conformational effects of site-directed mutagenesis of the lysine residues in the activation domain of human Pg and the consequences of these changes on the activation rate of Pg.

TECHNICAL SKILLS:

Experienced in protein expression using yeast, insect and E. coli systems.

Expert in protein purification using fast protein liquid chromatography (FPLC) with a variety of methods: affinity, metal chelation, ionic exchange, hydrophobic interactions, and gel filtration.

Mastered biochemical and biophysical characterization of proteins using isothermal calorimetry (ITC), differential scanning calorimetry (DSC), circular dichroism (CD), surface plasmon resonance (SPR), mass spectrometry (MALDI-TOF), and analytical ultracentrifugation (AU).

MANAGEMENT AND ORGANIZATIONAL SKILLS:

Trained several postdoctoral research associates, co-op, undergraduate and graduate students in the use of lab equipment and the implementation of various research techniques. Managed the projects of one co-op and one undergraduate student.

Contributed as a member of a team of four people to successfully move an entire laboratory consisting of 30 people and 7 separate rooms to a new location.

In charge of keeping inventory and ordering supplies and reagents for the entire laboratory. Used Sciquest ordering system.

EDUCATION

August 2008. Ph. D. in Biochemistry, University of Notre Dame. Notre Dame, IN.

March 2000. B.Sc. Biology, University of Los Andes, Mérida, Venezuela.

PROFESSIONAL TRAINING

November 2017. Certificate of Completion for Inform GMT 5.5 for Site Users (CRC) EDC System. Oracle.

October 2017. Certificate of Completion for Columbia Suicide Severity Rating Scale (C-SSRS) Initial Training V1.1 and Refresher Training V1.3. Visual Health Information/BlueCloud.

July-October 2017. Clinical Research Management Course. New York Academy of Sciences. 13-week online course that provides extensive practical training in clinical research and clinical data management including the use of Oracle Clinical Trial Management System.

September 2017. Certificate of Completion for Rave EDC and Rave Modules. Medidata Academy.

September 2017. Certificate of Completion for ACUVIA Electronic Regulatory Binder System (Complion).

September 2017. Certificate of Proficiency in the Appropriate Use of Clinical Looking Glass, a medical records analytical online tool. Montefiore Medical Center and Streamline Health.

August 2017. Certificate of Completion for the online training course for International Air Transport Association (IATA), Infectious Substances and Dry Ice (2017). INC Research.

August 2017. Certificate of Completion for the online training module of Good Clinical Practice ICH E6 revision 2, Changes for Site Personnel Working on Clinical Research Studies. INC Research.

August 2017. Certificate of Completion for the EPIC-REQ-View Only Ambulatory EpicCare eLearning online training course. Montefiore Medical Center.

June 2017. Certificate of Completion for Columbia Suicide Severity Rating Scale (C-SSRS) Screener Version 1. Visual Health Information/BlueCloud.

April 5 and 6, 2017. Clinical Research Investigator Training Program organized by Pfizer. Montefiore Medical Center, Bronx, NY.

Winter 2016-Spring 2017. Design and Conduct of Clinical Research. Nine-weeks seminar course. Albert Einstein College of Medicine, Bronx, NY.

December 2016. C14 Fitness certificate issued by the New York City Fire Department for supervising non-production chemical laboratories. Albert Einstein College of Medicine, Bronx, NY.

November 2016. Collaborative Institutional Training Initiative (CITI Program) certificates for Good Clinical Practice Course for Clinical Trials Involving Drugs (ICH focus) GCP-ICH, Biomedical Research (including EPI), Biomedical Research with Drugs/Devices and Albert Einstein College of Medicine Conflict of Interest (COI) Course.

November 2013. Training in Neurotherapeutics Discovery and Development for academic Scientists. Course supported by the National Institute of Neurological Disorders and Stroke and the NIH Blueprint for Neuroscience Research. Bethesda, MD.

TEACHING AND MENTORING EXPERIENCE

Lecturer in Immunology, Biotechnology and Pharmacogenomics, Gregory School of Pharmacy, Palm Beach Atlantic University, West Palm Beach, FL (2012)

Prepared and instructed lectures in Immunology during Spring 2012 semester for 50 students. Formulated questions for written exams and graded tests.

Teaching Assistant in Organic Chemistry Laboratory, Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN (2001- 2002)

Instructed, examined, supervised and tutored 40 students per semester.

Bioinformatics user support at the Supercomputing Center of the University of Los Andes (CeCalCULA), Mérida, Venezuela (1998- 2000)

Developed, wrote, translated into Spanish and published online manuals and documentation for the use of Bioinformatics software and databases for DNA and protein sequence analyses and for literature searches. Prepared, organized and taught workshops in the use of such Bioinformatics tools and databases.

Teaching Assistant in Biochemistry Laboratory, Biology Department, University of Los Andes, Mérida, Venezuela (1997- 2000)

Responsible for teaching introductory biochemistry laboratory class to 15-20 students per semester.

HONORS AND AWARDS

Selected to participate in the Neurotherapeutics Discovery and Development course for Academic Scientists, 2013.

Third place for oral presentation in the Chemistry-Biochemistry-Biology Interface retreat, 2008.

Magna Cum Laude distinction and Valedictorian at graduation ceremony, 2000.

Gold Medal "Luis Ribas Dávila", for the highest academic performance among all Biology Major students, 1998.

WRITING EXPERIENCE

2013-present contributing science writer at ASBMB Today, the monthly scientific news magazine of the American Society for Biochemistry and Molecular Biology (http://www.asbmb.org/asbmbtoday/).

REVIEWING EXPERIENCE

2013-present. Reviewer of research articles aspiring to appear in the Journal of Central Nervous System Disease, Biochemistry Insights, Drug Design, Development and Therapy and the Journal of Cell Death.

2014-present. Reviewer for the Graduate Women in Science National Fellowship applications.

POSTERS AND ORAL PRESENTATIONS

Poster #234.17: Neuroscience 2014, Society for Neuroscience, November 2014. Figuera-Losada M*., Stathis M., Wozniak K.M., Stockwell B., Rojas C, Slusher B.S. In vitro inhibition of glutamate/cystine transporter in microglial cells reduces excitotoxic conditions.

Poster #61: 12th International Symposium on NeuroVirology and 2013 Conference on HIV in the Nervous System, October 2013. Mariana Figuera-Losada*, Marigo Stathis, Camilo Rojas and Barbara S. Slusher. Development of microglial activation phenotypic assays as a tool to screen therapeutic agents for neuroAIDS and other neuroinflammatory disorders.

Poster #62: 12th International Symposium on NeuroVirology and 2013 Conference on HIV in the Nervous System, October 2013. Mariana Figuera-Losada*, Marigo Stathis, Norman J. Haughey, Camilo Rojas and Barbara S. Slusher. Screening for neutral sphingomyelinase inhibitors as a novel target in human immunodeficiency virus-1 associated neurocognitive disorders.

Poster: Gordon Research Conference Phosphorylation & G-Protein Mediated Signaling Networks, June 2012. Mariana Figuera-Losada* and Philip V. LoGrasso. Activity and substrate binding differences between cJun N-terminal kinase (JNK) variants.

Oral Presentation: Chemistry-Biochemistry-Biology Interface Retreat, University of Notre Dame, Indiana, USA, March 2008. Mariana Figuera-Losada* and F. J. Castellino. Human Plasminogen and Group A Streptococcal M-like Protein: Interaction, Conformational Transition and Activation.

Oral Presentation #09-04: XIXth International Congress on Fibrinolysis & Proteolysis, Vienna, July 2008. Mariana Figuera-Losada, Mary Prorok, Qihua Fu, Martina L. Sanderson-Smith, Mark J. Walker, Marie Ranson and Francis J. Castellino*. Human Plasminogen and Group A Streptococcal M-like Protein: Interaction, Conformational Transition and Activation.

Oral Presentation #298: The 18th International Congress on Fibrinolysis and Proteolysis: Proteolysis in the Postgenomic Era, California, USA August, 2006. M. Prorok*, S. Cnudde, M. Figuera, F.J. Castellino, and J.H. Geiger. The Effects of a PAM-derived Peptide on Angiostatin Structure and Zymogenicity of Plasminogen.

Poster #114: Xth International Workshop on Molecular & Cellular Biology of Plasminogen Activation. Washington DC, USA, 2005. M. Figuera* and F.J. Castellino. Identification of the Residues and Domains of Human Plasminogen Implicated in the Conformational Transition during Its Activation.

PUBLICATIONS

(1) Figuera-Losada, M.; Thomas, A. G.; Stathis, M.; Stockwell, B. R.; Rojas, C.; Slusher, B. S. Development of a primary microglia screening assay and its use to characterize inhibition of system xc- by erastin and its analogs. Biochemistry and Biophysics Reports 2017, 9, 266-272.

(2) Figuera-Losada, M.; Stathis, M.; Dorskind, J. M.; Thomas, A. G.; Bandaru, V. V.; Yoo, S. W.; Westwood, N. J.; Rogers, G. W.; McArthur, J. C.; Haughey, N. J.; Slusher, B. S.; Rojas, C. Cambinol, a novel inhibitor of neutral sphingomyelinase 2 shows neuroprotective properties. PloS one 2015, 10, e0124481.

(3) Potter, M. C.; Figuera-Losada, M.; Rojas, C.; Slusher, B. S. Targeting the glutamatergic system for the treatment of HIV-associated neurocognitive disorders. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology 2013, 8, 594-607.

(4) Figuera-Losada, M.; Rojas, C.; Slusher, B. S. Inhibition of Microglia Activation as a Phenotypic Assay in Early Drug Discovery. Journal of biomolecular screening 2013.

(5) Feng, Y.; Chambers, J. W.; Iqbal, S.; Koenig, M.; Park, H.; Cherry, L.; Hernandez, P.; Figuera-Losada, M.; Lograsso, P. V. A Small Molecule Bidentate-Binding Dual Inhibitor Probe of the LRRK2 and JNK Kinases. ACS Chem Biol 2013.

(6) Laughlin, J. D.; Nwachukwu, J. C.; Figuera-Losada, M.; Cherry, L.; Nettles, K. W.; LoGrasso, P. V. Structural mechanisms of allostery and autoinhibition in JNK family kinases. Structure 2012, 20, 2174-2184.

(7) Figuera-Losada, M.; LoGrasso, P. V. Enzyme kinetics and interaction studies for human JNK1beta1 and substrates activating transcription factor 2 (ATF2) and c-Jun N-terminal kinase (c-Jun). J Biol Chem 2012, 287, 132**-*****.

(8) Chambers, J. W.; Cherry, L.; Laughlin, J. D.; Figuera-Losada, M.; Lograsso, P. V. Selective inhibition of mitochondrial JNK signaling achieved using peptide mimicry of the Sab kinase interacting motif-1 (KIM1). ACS Chem Biol 2011, 6, 808-818.

(9) Kamenecka, T.; Jiang, R.; Song, X.; Duckett, D.; Chen, W.; Ling, Y. Y.; Habel, J.; Laughlin, J. D.; Chambers, J.; Figuera-Losada, M.; Cameron, M. D.; Lin, L.; Ruiz, C. H.; LoGrasso, P. V. Synthesis, biological evaluation, X-ray structure, and pharmacokinetics of aminopyrimidine c-jun-N-terminal kinase (JNK) inhibitors. J Med Chem 2010, 53, 419-431.

(10) Figuera-Losada, M.; Ranson, M.; Sanderson-Smith, M. L.; Walker, M. J.; Castellino, F. J.; Prorok, M. Effects on human plasminogen conformation and activation rate caused by interaction with VEK-30, a peptide derived from the group A streptococcal M-like protein (PAM). Biochim Biophys Acta 2010, 1804, 1342-1349.

(11) Fu, Q.; Figuera-Losada, M.; Ploplis, V. A.; Cnudde, S.; Geiger, J. H.; Prorok, M.; Castellino, F. J. The lack of binding of VEK-30, an internal peptide from the group A streptococcal M-like protein, PAM, to murine plasminogen is due to two amino acid replacements in the plasminogen kringle-2 domain. J Biol Chem 2008, 283, 1580-1587.

(12) Figuera-Losada, M. P., M.; Fu, Q.; Sanderson-Smith, M. L.; Walker, M. J.; Ranson, M.; Castellino, F.J. In Tilte, Vienna, Austria2008.

(13) Prorok, M. C., S.; Figuera, M.; Castellino, F.J.; Geiger, J.H. In Tilte, California, USA2006.

(14) Prorok, M., Figuera, M., Cnudde, S., Geiger, J., & Castellino, F. . The Effects of a PAM-derived Peptide on the Structure and Zymogenicity of Plasminogen. JOURNAL OF THROMBOSIS AND HAEMOSTASIS 2006, 4, 183.

(15) Mundada, L. V.; Prorok, M.; DeFord, M. E.; Figuera, M.; Castellino, F. J.; Fay, W. P. Structure-function analysis of the streptokinase amino terminus (residues 1-59). J Biol Chem 2003, 278, 244**-*****.

(16) Iwaki, T.; Figuera, M.; Ploplis, V. A.; Castellino, F. J. Rapid selection of Drosophila S2 cells with the puromycin resistance gene. Biotechniques 2003, 35, 482-484, 486.

(17) Avilan, L.; Calcagno, M.; Figuera, M.; Lemus, L.; Puig, J.; Rodriguez, A. M. Interaction of Leishmania mexicana promastigotes with the plasminogen-plasmin system. Mol Biochem Parasitol 2000, 110, 183-193.

SOCIETY MEMBERSHIPS

2017-Present International Parkinson and Movement Disorder Society

2017-present New York Academy of Sciences

2014-present Sigma Xi The Scientific Research Society

2013-present Graduate Women in Science. Omicron Chapter

_ENREF_12012-present. Biochemical Society

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