UH3CA256967
Cooperative Agreement
Overview
Grant Description
A Streamlined Platform for Phosphoproteome Mapping of Human Tissues - Abstract
Recent technological advances in genomics, transcriptomics, and proteomics allow for rapid generation of comprehensive 3D-human tissue maps for biomolecules DNAs, RNAs, and proteins at the single-cell resolution in the HuBMAP Consortium. However, single-cell technologies for characterizing functional modifications are lagging far behind but equally important as these existing omics technologies.
Protein phosphorylation is one of the most important modifications and often used as an indicator of signaling pathway activation (cell functional state). The lack of high-spatial-resolution phosphoproteomic characterization of human tissues in the HuBMAP Consortium represents a significant knowledge gap for achieving a more complete understanding of how tissue heterogeneity impacts human health.
The objective of this TTD application is to address this gap by developing a convenient streamlined platform for enabling automated high-resolution 3D-phosphoproteome mapping of human tissues. The project feasibility is strongly supported by our recent progress in many aspects of technology development:
1) Carrier-Assisted Sample Preparation (CASP) for both global and targeted proteomics analysis of 1-100 cells;
2) A Boosting to Amplify Signal with Isobaric Labeling (BASIL) strategy for high-throughput single-cell proteomics;
3) BASIL/TIP-IMAC (Immobilized Metal Affinity Chromatography) for rapid phosphoproteomic analysis of small numbers of cells;
4) Advanced liquid chromatography (LC) separation and disruptive mass spectrometry (MS) technologies (e.g., multi-emitter array technology and sub-ambient pressure ionization with nanoelectrospray (SPIN) source) for improving MS detection sensitivity.
In the UG3 phase, Aim 1 will focus on the development of a streamlined platform through 1) improving phospho-recovery by developing an improved CASP/online IMAC platform for automated processing and phospho-enrichment, and 2) leveraging multiple disruptive technologies developed at our group with integration of a high-efficiency multi-emitter SPIN (MSPIN) source and BASIL-based sample multiplexing for significantly improving MS sensitivity by ~50-fold and sample throughput by >20-fold. The streamlined platform will allow for precise quantification of ~1,000 phosphosites in single cells and ~7,000 phosphosites in 10 cells with >1,000 samples per day.
Aim 2 will demonstrate the streamlined platform for enabling 2D-phosphoproteome mapping of mouse uterine tissues when combined with laser capture microdissection (LCM) and standard tube-based voxel collection.
In the UH3 phase (Aim 3), we will further optimize the streamlined platform for automated robust phosphoproteomic analysis of LCM-dissected human tissue sections. We then will validate the streamlined platform for high-resolution 3D-phosphoproteome mapping of human breast and uterine tissues and other human tissues from the HuBMAP Consortium. An easy-to-use visualization tool will be developed to generate 3D maps that can be quickly and easily accessible by the research community. With its antibody-free feature, the streamlined platform can be equally applicable to any types of tissues.
We envision that the streamlined platform will become an indispensable tool for high-resolution 3D-phosphoproteome mapping of human tissues in the HuBMAP Consortium and extend the HuBMAP toolbox for 3D-mapping of functional modifications. In turn, it will make substantial contributions to improve our understanding of tissue biology and accelerate the movement toward precision medicine.
Recent technological advances in genomics, transcriptomics, and proteomics allow for rapid generation of comprehensive 3D-human tissue maps for biomolecules DNAs, RNAs, and proteins at the single-cell resolution in the HuBMAP Consortium. However, single-cell technologies for characterizing functional modifications are lagging far behind but equally important as these existing omics technologies.
Protein phosphorylation is one of the most important modifications and often used as an indicator of signaling pathway activation (cell functional state). The lack of high-spatial-resolution phosphoproteomic characterization of human tissues in the HuBMAP Consortium represents a significant knowledge gap for achieving a more complete understanding of how tissue heterogeneity impacts human health.
The objective of this TTD application is to address this gap by developing a convenient streamlined platform for enabling automated high-resolution 3D-phosphoproteome mapping of human tissues. The project feasibility is strongly supported by our recent progress in many aspects of technology development:
1) Carrier-Assisted Sample Preparation (CASP) for both global and targeted proteomics analysis of 1-100 cells;
2) A Boosting to Amplify Signal with Isobaric Labeling (BASIL) strategy for high-throughput single-cell proteomics;
3) BASIL/TIP-IMAC (Immobilized Metal Affinity Chromatography) for rapid phosphoproteomic analysis of small numbers of cells;
4) Advanced liquid chromatography (LC) separation and disruptive mass spectrometry (MS) technologies (e.g., multi-emitter array technology and sub-ambient pressure ionization with nanoelectrospray (SPIN) source) for improving MS detection sensitivity.
In the UG3 phase, Aim 1 will focus on the development of a streamlined platform through 1) improving phospho-recovery by developing an improved CASP/online IMAC platform for automated processing and phospho-enrichment, and 2) leveraging multiple disruptive technologies developed at our group with integration of a high-efficiency multi-emitter SPIN (MSPIN) source and BASIL-based sample multiplexing for significantly improving MS sensitivity by ~50-fold and sample throughput by >20-fold. The streamlined platform will allow for precise quantification of ~1,000 phosphosites in single cells and ~7,000 phosphosites in 10 cells with >1,000 samples per day.
Aim 2 will demonstrate the streamlined platform for enabling 2D-phosphoproteome mapping of mouse uterine tissues when combined with laser capture microdissection (LCM) and standard tube-based voxel collection.
In the UH3 phase (Aim 3), we will further optimize the streamlined platform for automated robust phosphoproteomic analysis of LCM-dissected human tissue sections. We then will validate the streamlined platform for high-resolution 3D-phosphoproteome mapping of human breast and uterine tissues and other human tissues from the HuBMAP Consortium. An easy-to-use visualization tool will be developed to generate 3D maps that can be quickly and easily accessible by the research community. With its antibody-free feature, the streamlined platform can be equally applicable to any types of tissues.
We envision that the streamlined platform will become an indispensable tool for high-resolution 3D-phosphoproteome mapping of human tissues in the HuBMAP Consortium and extend the HuBMAP toolbox for 3D-mapping of functional modifications. In turn, it will make substantial contributions to improve our understanding of tissue biology and accelerate the movement toward precision medicine.
Awardee
Funding Goals
NOT APPLICABLE
Grant Program (CFDA)
Awarding Agency
Place of Performance
Washington
United States
Geographic Scope
State-Wide
Related Opportunity
Analysis Notes
Amendment Since initial award the End Date has been extended from 08/31/24 to 08/31/25 and the total obligations have increased 100% from $600,000 to $1,200,000.
Battelle Memorial Institute was awarded
A streamlined platform for phosphoproteome mapping of human tissues
Cooperative Agreement UH3CA256967
worth $1,200,000
from the National Institute of Allergy and Infectious Diseases in September 2020 with work to be completed primarily in Washington United States.
The grant
has a duration of 5 years and
was awarded through assistance program 93.310 Trans-NIH Research Support.
The Cooperative Agreement was awarded through grant opportunity Transformative Technology Development for the Human BioMolecular Atlas Program (UG3/UH3 Clinical Trial Not Allowed).
Status
(Ongoing)
Last Modified 1/21/25
Period of Performance
9/9/20
Start Date
8/31/25
End Date
Funding Split
$1.2M
Federal Obligation
$0.0
Non-Federal Obligation
$1.2M
Total Obligated
Activity Timeline
Transaction History
Modifications to UH3CA256967
Additional Detail
Award ID FAIN
UH3CA256967
SAI Number
UH3CA256967-2638069157
Award ID URI
SAI UNAVAILABLE
Awardee Classifications
Nonprofit With 501(c)(3) IRS Status (Other Than An Institution Of Higher Education)
Awarding Office
75NC00 NIH NATIONAL CANCER INSTITUTE
Funding Office
75NA00 NIH OFFICE OF THE DIRECTOR
Awardee UEI
CWKJEXDG79A7
Awardee CAGE
1A453
Performance District
WA-90
Senators
Maria Cantwell
Patty Murray
Patty Murray
Budget Funding
| Federal Account | Budget Subfunction | Object Class | Total | Percentage |
|---|---|---|---|---|
| Office of the Director, National Institutes of Health, Health and Human Services (075-0846) | Health research and training | Grants, subsidies, and contributions (41.0) | $1,200,000 | 100% |
Modified: 1/21/25