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From Single Cells to Tissues: a Novel Mass Spectrometry Approach for Bioanalysis

Award Number: R01GM116116
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/01/2015
PERIOD OF PERFORMANCE END DATE: 07/31/2021

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From Single Cells to Tissues: a Novel Mass Spectrometry Approach for Bioanalysis - DESCRIPTION (provided by applicant): There is a fundamental lack of understanding in biology on the single cell level, especially in human disease biology. This gap in knowledge encompasses both how individual cells function and also how single cells combine and collaborate to construct multi-cellular creations, such as how many cancer cells associate to form a tumor. Traditionally, bioanalysis has largely been conducted through large numbers (i.e. thousands or millions) of cells being destroyed and mixed together to form a completely artificial construct: the cell lysate. Cell lysates are prepared from a large number of heterogeneous cells, either present in tissue samples or grown in culture. Bioanalysis of lysates are an amalgamated average of many heterogeneous cells subjected to artificial sample preparation, and therefore, standard bioanalytical methods have been largely unable to define single cell biology. Recently, single cell analysis (SCA) has become possible through advancements in many experimental methods. Due to its unparalleled detection sensitivity, mass spectrometry (MS) is one of the most promising analytical methods to be deployed for single cell analysis. Single-cell mass spectrometry (SCMS) and mass spectrometry imaging (MSI) are emerging techniques that could potentially revolutionize bioanalytical science in basic scientific and biomedical research. We have developed a miniaturized sampling and ionization device, the Single-probe, capable of performing single cell mass spectrometry (SCMS) in real time under ambient conditions. The Single-probe, with a sampling tip smaller than eukaryotic cells (<10 µm), can be inserted into individual living cells to sample the intracellular compounds for immediate MS analysis. The Single-probe SCMS technique has produced highly sensitive detection of intracellular compounds, including cell metabolites, lipids and anticancer compounds, inside living single cells. The Single-probe has also been used successfully in MSI experiments to reconstruct the spatial distribution of species (e.g. lipids and cell metabolites) from sections of animal tissue. Our long-term research goal is to fully establish the Single- probe based techniques in SCMS and MSI for broad utilization in bioanalysis. The objective of this application is to develop the Single-probe MS technology as a first-in-class tool for quantitative SCMS (qSCMS) in cells and for MSI-based investigation of tumors. This research objective will be accomplished through completing the following three Specific Aims: 1) create quantitative single cell mass spectrometry (qSCMS) for compounds inside of single cells from cultured cell lines; 2) use the Single-probe MS to analyze single cells in in vitro tumors; and, 3) perform Single-probe MSI measurements of the in vitro and animal-derived tumors. The Single-probe MS is an innovative technology in that it provides new experimental capabilities for single cell analysis and tissue imaging. The proposed research is significant because the Single-probe MS experimental applications (i.e. qSCMS and MSI), once developed, will allow new understandings in single cell and tumor biology that are not currently possible.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2023 ( Subtotal = $8,534 )
2023	2019	UNIVERSITY OF OKLAHOMA	660 PARRINGTON OVAL RM 301	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	000	5	11/23/2022	NON-COMPETING CONTINUATION	$8,534
														Subtotal = $8,534

Issue Date FY: 2022 ( Subtotal = -$240,466 )
2022	2019	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	000	5	3/17/2022	NON-COMPETING CONTINUATION	-$240,466
														Subtotal = -$240,466

Issue Date FY: 2019 ( Subtotal = $297,005 )
2019	2019	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	000	5	8/1/2019	NON-COMPETING CONTINUATION	$297,005
														Subtotal = $297,005

Issue Date FY: 2018 ( Subtotal = $318,606 )
2018	2018	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	000	4	8/21/2018	NON-COMPETING CONTINUATION	$318,606
														Subtotal = $318,606

Issue Date FY: 2017 ( Subtotal = $317,268 )
2017	2017	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	003	3	8/31/2017	SUPPLEMENT FOR EXPANSION	$20,080
2017	2017	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	002	3	7/24/2017	NON-COMPETING CONTINUATION	$297,188
2017	2016	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	001	2	2/16/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$45,019
2017	2015	UNIVERSITY OF OKLAHOMA	1000 ASP AVE RM 105	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	000	1	2/16/2017	NEW	-$45,019
														Subtotal = $317,268

Issue Date FY: 2016 ( Subtotal = $297,276 )
2016	2016	UNIVERSITY OF OKLAHOMA	201 STEPHENSON PARKWAY	NORMAN	OK	73019	CLEVELAND	USA	Biomedical Research and Research Training	000	2	7/19/2016	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$297,276
														Subtotal = $297,276

Issue Date FY: 2015 ( Subtotal = $297,361 )
2015	2015	UNIVERSITY OF OKLAHOMA NORMAN CAMPUS	1000 ASP AVENUE	NORMAN	OK	73109		USA	Biomedical Research and Research Training	000	1	7/27/2015	NEW	$297,361
														Subtotal = $297,361

Grand Total All Awards = $1,295,584

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From Single Cells to Tissues: a Novel Mass Spectrometry Approach for Bioanalysis

Award Number: R01GM116116

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 08/01/2015

PERIOD OF PERFORMANCE END DATE: 07/31/2021

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