# Core B: Mass spectrometry, proteomics, metabolomics and lipidomics

> **NIH NIH P01** · BRIGHAM AND WOMEN'S HOSPITAL · 2022 · $170,592

## Abstract

Program Director/Principal Investigator (Last, First, Middle): Kwiatkowski, David, J. et al., Core B; Asara
Project Summary/Abstract
The mass spectrometry core has expertise in proteomics/phosphoproteomics, metabolomics and lipidomics
resources to enable the three major P01 projects achieve success in uncovering the molecular mechanisms of
Hamartoma syndromes and related cancers in the TSC1-TSC2 pathways for new drug targets and novel
therapies using tandem mass spectrometry (LC-MS/MS). The core utilizes both high resolution hybrid Orbitrap
(QExactive) mass spectrometry and hybrid triple quadrupole (QTRAP) mass spectrometry. For proteomics,
microcapillary tandem mass spectrometry (LC-MS/MS) services will include protein complex identification, post-
translational modification (PTM) site mapping such as phosphorylation, ubiquitination, acetylation, etc. and the
relative and absolute quantification of peptides/proteins using both stable isotope labeling (SILAC and TMT) and
label-free quantification [spectral counting, total ion current (TIC), multiple reaction monitoring (MRM)]. These
studies will be performed from cell lines, xenografts in addition to in vivo tissue sources from mice and human
tumors. We have developed expertise in metabolomics profiling and services will include polar metabolite
profiling using selected reaction monitoring (SRM) with polarity switching to target more than 300 molecules in
15 min. We will profile cells, tumor tissues and biological fluids using both steady-state profiling and 13C and 15N
stable isotope labeled flux experiments to determine which metabolic pathways are altered in cells harboring
defects in the TSC1/2 related pathways. Non-targeted metabolomic profiling will also be performed to discover
novel metabolic targets. Core B has recently developed a non-targeted lipidomics platform based on high
resolution mass spectrometry with polarity switching with novel software to identify more than 1000 lipid ions
(phospholipids, triglycerides, free fatty acids, etc.) in less than 30 min. using reversed-phase LC-MS/MS. We will
also develop stable isotope flux for lipidomics experiments. In addition to running samples for Projects 1-3, Core
B has also developed a serial-omics technology that utilizes the preparation of a single tumor, cell or bodily fluid
sample for performing three different –omics (global phosphoproteomics/proteomics, metabolomics and
lipidomics) via partitioning liquid-liquid extraction layers. We will also continue to develop -omics strategies to
overlap model species (drosophila) to cancer cells and tumor tissue to uncover conserved biological interactions
for potential biomarker targets in TSC1/2 and related pathways.

## Key facts

- **NIH application ID:** 10460144
- **Project number:** 5P01CA120964-15
- **Recipient organization:** BRIGHAM AND WOMEN'S HOSPITAL
- **Principal Investigator:** John M Asara
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $170,592
- **Award type:** 5
- **Project period:** 2006-12-01 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10460144

## Citation

> US National Institutes of Health, RePORTER application 10460144, Core B: Mass spectrometry, proteomics, metabolomics and lipidomics (5P01CA120964-15). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10460144. Licensed CC0.

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