You are here

Tory Schaaf, Postdoctoral Associate

Tory I am directly involved in the development of novel fluorescence tools for the discovery of cardiac calcium pump therapeutics. The sarco/endoplasmic reticulum calcium ATPase (SERCA) is the calcium pump responsible for maintaining cellular calcium homeostasis. Diminished SERCA function has been directly linked to numerous degenerative disease states, such as heart failure. The pathological progression of heart failure is associated with an elevated level of cytosolic calcium, and impairs the function of the muscle contraction-cycle. The overarching goal of my research is to discover novel small-molecule effectors, capable of enhancing SERCA’s ability to pump and store calcium within the sarcoplasmic reticulum (SR). Drugs that increase the calcium pumping efficiency of SERCA will restore calcium homeostasis by reducing the calcium content in the cytosol, and enhance impaired cardiac function.

The process of drug discovery is a high-risk effort, and involves screening millions of small-molecules to fortuitously discover a lead compound with high-therapeutic potential. The precise placement of two fluorescent proteins at specific locations along SERCA’s cytosolic headpiece, allows for the detection of fluorescence resonance energy transfer (FRET) between donor and acceptor fluorescent proteins. I have generated human cell lines that overexpress this fluorescent fusion protein and created a live-cell biosensor. The rate of energy transfer (FRET) is dependent on the distance between the fluorescent probes and linked to the enzymatic activity of SERCA. FRET tracks SERCA’s structural status, while it pumps calcium into the sarcoplasmic reticulum. These biosensors are grown in vast quantities, harvested, and utilized for high-throughput drug screening. The cells are dispensed into high-density microplates, where each well contains a different compound.

Through my research, I have demonstrated that FRET can be utilized for the purposes of high-throughput drug screening by using proprietary fluorescence technology, capable of recording the nanosecond fluorescence decay rate (lifetime) and the full emission spectrum. Both lifetime and spectral modes offer incredibly fast speeds, with high resolution and precision. High-throughput screening by lifetime mode offers the advantage of resolving the structural status of the FRET biosensor because the mole fraction of each structural state is assessed, and candidate compounds found during the screening process can be characterized by their structural effect on the biosensor. High-throughput screening by spectral mode increases assays precision by taking into account the shape of the fluorescence emission spectrum. The shapes of these spectra are decomposed into the contribution of known components by a novel spectral unmixing method, and further used to accurately evaluate FRET. When coupled with lifetime mode, spectral-based drug screening increases assay precision and removes artifacts from cellular autofluorescence and fluorescent compounds. The complementary advantages of coupling spectral and lifetime fluorescence measurements significantly reduce the rate of false-positives from high-throughput drug screens.  The development of these technologies and FRET biosensor assays, drastically increases the probability of identifying a novel drug with great therapeutic potential.

Example of spectral unmixing method used to determine FRET from a live-cell biosensor

 

I am currently interested in applying these novel technologies for the development of new FRET-biosensors targeting other promising therapeutic pathways. I have training and research experience in a broad range of fields (neuroscience, immunology, stem cells, molecular biology, structural biology, and optical fluorescence). My long-term career goal is to work in industry but still remain in close contact with academic research to hopefully make a lasting impact in both areas.

 

EDUCATION

2012-2017    

University of Minnesota, Minneapolis, MN                                   Ph.D., Biochemistry, Molecular Biology, and Biophysics           Advisor: David. D. Thomas, PhD

 2007-2009

University of Minnesota, Minneapolis, MN                                   BS,Neuroscience                                                                                 BS, Biochemistry

2005-2007

University of Wisconsin River Falls, WI

 

 

 

 

 

 

 

        

 

 

 

TEACHING & RESEARCH EXPERIENCE

2018HHMI BioInteractive Training
2018Preparing Future Faculty Certificate, University of Minnesota
2018 SummerBIOL 1100, Intro to Genetics (TREM Scholar at Normandale Community College)
2015 Summer BIOC 4125, Laboratory in Molecular Biology and Biotechnology
2014 FallBIOC 4125, Laboratory in Molecular Biology and Biotechnology
2008-2009 Undergraduate Research Assistant, University of Minnesota (Mark Thomas, PhD)

 

PROFESSIONAL EXPERIENCE

 

LinkedIn Profile

2017 – Present                       Post-Doctoral Fellow, University of Minnesota, Minneapolis, MN
Perform research and teach undergraduate level biology courses. Publish scientific articles and write research grants to secure funding for lab projects, collaborative projects, and industrial focused research. Prepare tutorials, protocols, and help documents. Assist, train, and mentor high school, undergraduate, and graduate students from multiple departments at the UMN and local community colleges. Research mentor for underrepresented students from local community colleges. Assist and collaborate with UMN researchers to develop new drug discovery assays and technologies. Present research at international conferences. Publish and patent research methods and protocols.
 
2018 - Present                      High-throughput Screening (HTS) Manager, Biophysical Technology Core, University of Minnesota, Minneapolis, MN
Oversee operation of fluorescence instrumentation, Train fluorescence instrument users, Coordinate service and maintenance of instrumentation, Consult on the development of novel FRET-based screening assays for large-scale drug discovery efforts, Participate in multiple weekly meetings to support operations at UMN core facilities, Develop standard operating procedures for internal and external users, Perform quality control experiments to optimize performance of instrumentation and data analysis practices.
 
2016-2018                            Scientist, Fluorescence Innovations Inc., Minneapolis, MN
Part time position at Fluorescence Innovations, a small company that develops and builds novel fluorescence plate reader technology. My role was to evaluate their new technologies, optimize novel screening assays, and coordinate large group efforts for a NIH-sponsored STTR phase II grant. Consulted on new projects to help gauge the likelihood of success, decide project tangibility, and aid in experiment design. These projects require a high-level of coordination between cell biologists and engineers. I was also involved in the training of techniques, concepts, data acquisition, and data analysis. Other responsibilities included maintaining the company budget, preparing weekly progress reports, writing grants, and effectively communicating ideas across groups of professors at the University of Minnesota.
 
2011-2013                            Research Associate, R&D Systems, Minneapolis MN.
Performed tissue culture; Transfected expression constructs for stable clone development. Screened for recombinant protein expression utilizing western blot, fluorescence microscopy, and flow cytometry; Utilized DNA cloning techniques for gene-isolation and VersaClone cDNA vectors; Effectively planned, organized, and multi-tasked to complete assignments in an efficient manner; Interacted positively with flow cytometry, cell culture, protein purification, antibody, and assay development group; Demonstrated ability to work independently and with teams to complete objectives and goals.
 
2009-2011                            Junior Scientist, University of Minnesota. Minneapolis. MN. Laboratory of Dr. Rita Perlingeiro. Lillehei Heart Institute
Performed gene therapy preclinical trials; Expressed micro-utrophin in human or mouse induced pluripotent stem cells lacking the dystrophin gene, which were then differentiated into skeletal muscle myoblasts and transplanted into animal models of muscular dystrophy; Performed molecular biology to generate new cell lines with cre-recombinase, retroviral, lentiviral, and sleeping beauty transposon systems; Performed RNA and protein analyses utilizing qRT-PCR and western blotting; Problem solved and developed new protocols for lentiviral production and cell sorting using FACS; Presented data and results bi-weekly to large groups; Aided in animal husbandry for an SPF animal colony with over 1,500 animals.
 
2009                                      Junior Scientist, University of Minnesota. Minneapolis. MN. Laboratory of Dr. Keli Hippen. Dept. of Bone Marrow Transplantation.
Performed preclinical trials in the involving the suppression of Graft-vs-host disease (GVHD) by regulatory T-cell transplantation; Performed flow cytometry and immunohistochemistry to document disease progression and treatment in xenogeneic model of GVHD; Performed large-scale cell culture experiments; Assessed and graded severity of disease using H+E staining and imaging software.; Isolated regulatory T-cells from peripheral blood mononuclear cells; Optimized multiplexed FACS analysis using up to eight different antibodies.

 

FUNDING & AWARDS

2017-Present         NIH K12 Training Research Educators in Minnesota Postdoctoral Scholar

2017                       NIH T32 Minnesota Muscle Postdoctoral Fellow 

2016                       Ross A. Gortner Award

2014-2016             NIH Chemistry-Biology Interface Training Grant

2012-2016             3M Graduate Fellowship

2013-2017             Arnold H. Johnson Graduate Fellowship

2014                       Charles Carr / William Peterson Award

2008-2009             Undergraduate Research Grant (UROP)

 

PATENTS

Tory Schaaf, Kurt Peterson, Benjamin Grant, David Thomas, and Greg Gillispie. 2017. High Precision FRET Assays. U.S. Patent Application 62450188. Filed on Jan. 25, 2017, Patent Pending.

PUBLICATIONS

Stroik D, S.Yuen, K Janicek, T Schaaf, J Li, D Ceholski, R Hajjar, R Cornea, and D Thomas. Targeting protein-protein interactions for therapeutic discovery via FRET-based high-throughput screening in living cells. Nature Scientific Reports. Accepted.

Lo, CH, N. Vunnam, AK Lewis, TL Chiu, BE Brummel, TM Schaaf, BD Grant, P Bawaskar, DD Thomas, and JN Sachs. An innovative high-throughput screening approach for discovery of small molecules that inhibit TNF Receptors. SLAS Discovery, Published online May 22, 2017.

Schaaf, TM, KC Peterson, BD Grant, P Bawaskar, S Yuen, J Li, JM Muretta, GD Gillispie, and DD Thomas. 2017. High-throughput spectral and lifetime-based FRET screening in living cells to identify small-molecule effectors of SERCA.  SLAS Discovery  22:262-273. PMC5323330.

Schaaf, TM, KC Peterson, BD Grant, DD Thomas, and GD Gillispie. 2017. Spectral unmixing plate reader: high-throughput, high-precision FRET assays in living cells. SLAS Discovery 22:250-261. PMC5323330.

Gruber, SJ, RL Cornea, J Li, KC Peterson, TM Schaaf, GD Gillispie, R Dahl, KM Zsebo, SL Robia, and DD Thomas. 2014. Discovery of enzyme modulators via high-throughput time-resolved FRET in living cells. J Biomol Screening 19:215-222 (cover article). PMC4013825.

Filareto, A, S Parker, R Darabi, L Borges, M Iacovino, T Schaaf, T Mayerhofer, J Chamberlain, J Ervasti, R McIvor, M Kyba, and R Perlingeiro. 2013. An ex vivo gene therapy approach to treat muscular dystrophy using inducible pluripotent stem cells. Nature Communications 4: 1549. PMC3595133.

 

ORAL PRESENTATIONS

Tory M. Schaaf, KC Peterson, BD Grant, J Li, P. Bawaskar, S Yuen, GD Gillispie, and DD Thomas. 2017. Spectral unmixing plate reader: High-throughput FRET-based screening in living cells to identify small-molecule effectors of SERCA. Biophysical Society 61th Annual Meeting, February 14, New Orleans, Louisiana.

Tory M. Schaaf, Ji Li, Samantha L. Yuen, Prachi Bawaskar, Benjamin D. Grant, Kurt C. Peterson, David D. Thomas, Gregory D. Gillispie. 2016. SUPR: Spectral unmixing plate reader for live-cell FRET biosensor drug screening. Biophysical Society 60th Annual Meeting, March 2, Los Angeles, California.

Tory M. Schaaf, Ji Li, Rocio Foncea, Simon J Gruber, Kurt C Peterson, Karl J Petersen, David A Bernlohr, Gregory D. Gillispie, and David D. Thomas. 2015. A two-color non-muscle SERCA FRET sensor for diabetes drug discovery using fluorescence lifetime detection. Chemistry-Biology Interface Career Development Workshop. August 10, Vanderbilt University, Tennessee.

Tory M. Schaaf, Ji Li, Rocio Foncea, Simon J Gruber, Kurt C Peterson, Karl J Petersen, David A Bernlohr, Gregory D. Gillispie, and David D. Thomas. 2015. A two-color non-muscle SERCA FRET sensor for diabetes drug discovery using fluorescence lifetime detection. Biophysical Society 59th Annual Meeting, February 10, Baltimore, Maryland.

POSTER PRESENTATIONS

Tory M. Schaaf, Ang Li, Benjamin D. Grant, Prachi Bawaskar, Evan Kleinboehl, Ji Li, Gregory D. Gillispie and David D. Thomas. Red-shifted FRET biosensors for high-throughput lifetime screening. Institutional Research and Academic Career Development Awards (IRACDA) 2018 NIH K12 Conference. July 16, 2018, Atlanta, Georgia. (Outstanding Poster Award).

Tory M. Schaaf, Ang Li, Benjamin D. Grant, Prachi Bawaskar, Evan Kleinboehl, Ji Li, Gregory D. Gillispie and David D. Thomas. Red-shifted fluorescent proteins improve FRET biosensors for high-throughput fluorescence lifetime screening. Biophysical Society 62th Annual Meeting, Feb 19, 2018, San Francisco, California.

Tory M. Schaaf, Kurt C. Peterson, Benjamin D. Grant, David D. Thomas, and Gregory D. Gillispie 2016. Spectral unmixing plate reader for high-precision FRET assays in living cells. Society of Laboratory Automation and Screening 5th Annual Meeting, January 23rd-27th. San Diego, California.

Tory M. Schaaf, Simon J Gruber, Kurt C Peterson, Karl J Petersen, Rocio Foncea, David A Bernlohr, Gregory D. Gillispie, and David D. Thomas. 2014. A two-color non-muscle SERCA FRET sensor for diabetes drug discovery using fluorescence lifetime detection. Biophysical Society 58th Annual Meeting, February 15th-19th, San Francisco, California.