# Core B: Mass Spectrometry, proteomics, metabolmics and lipidomics

> **NIH NIH P01** · BRIGHAM AND WOMEN'S HOSPITAL · 2024 · $142,417

## Abstract

Core B: 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
(Exploris 480, QExactive HF) mass spectrometry and hybrid triple quadrupole (QTRAP 6500/5500) mass
spectrometry. For proteomics, microcapillary tandem mass spectrometry (LC-MS/MS) services will include
protein complex identification, global 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 mouse/human tumors and drosophila models. 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 by HR-LC-MS/MS will also be performed to discover novel
metabolic targets. Core B will use non-targeted lipidomics based on high resolution mass spectrometry with
polarity switching with novel software to identify more than 1500 lipid ions (phospholipids, triglycerides, free fatty
acids, etc.) in less than 30 min. using reversed-phase LC-MS/MS. We will also use recently developed stable
13C/15N/18O isotope flux for lipidomics. In addition to running samples for Projects 1-3, Core B has developed
a serial-omics technology that utilizes the preparation of a single tisue, 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:** 10931472
- **Project number:** 5P01CA120964-17
- **Recipient organization:** BRIGHAM AND WOMEN'S HOSPITAL
- **Principal Investigator:** David J. Kwiatkowski
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $142,417
- **Award type:** 5
- **Project period:** 2007-04-24 → 2028-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10931472

## Citation

> US National Institutes of Health, RePORTER application 10931472, Core B: Mass Spectrometry, proteomics, metabolmics and lipidomics (5P01CA120964-17). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10931472. Licensed CC0.

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