ABSTRACT
This application is in response to NOT-GM-20-013 Notice of Special Interest (NOSI), Availability of
Administrative Supplements for Equipment Purchases for NIGMS Awardees for the parent grant 1R01
GM136874-01 entitled “Regenerative Lipid Mediators for the Management of Severe Burn Wounds.” In this
parent grant, we need to determine and optimize the sustained release of regenerative lipid mediators
(ReLiMs) from ReLiM loaded amino acid-based poly(ester amide) (AA-PEA) hydrogels (Agels) in Aim 1 and
from constructs that integrate adipose-tissue derived microvascular fragments (MVFs), ReLiMs, and Agel in
Aim 2 with the overall goal of optimal healing of burn wounds. Since the submission of the parent project,
samples requiring analysis by the current mass spectrometry (MS) instrumentation in our center’s Lipidomics
Core has markedly increased due to the increased research activities on lipids. Consequently, the samples for
this parent grant now greatly exceed the MS capacity of the Core. Moreover, we recently found that ReLiMs
are bioactive even at an unexpectedly low dose below the detection limit of the Core instruments. This means
the period for releasing bioactive ReLiM levels from optimal Agel dressings should be much longer than that
originally expected in the parent project, and we could avoid the Agel replacement on wounds and associated
complications. Thus, measuring time courses of ReLiM release from various Agels at low but bioactive doses
is vital. The ultra-high sensitive and fast hybrid triple quad-ion trap (QTRAP) 6500+MS is fully capable of such
measurement. Currently, no facility within a few hours’ drive has a QTRAP6500+MS system, so we have no
ready access to this equipment. Outsourcing our samples is also not reliable, time-effective, or affordable due
to their huge number and complexity. We have to conduct the time-consuming development of the LC-
QTRAP6500+ analysis methods tailored for each type of ReLiM-Agel and ReLiM-MvF-Agel. The maximum
funding provided by this supplement will be sufficient to purchase the MS part of the LC-QTRAP6500+MS
system, which is critical for successful implementation of the parent project. Our center will provide the
remaining components: an HPLC, a UV(/VIS) detector, and an autosampler compatible with the QTRAP6500+
MS, to form an autosampler-LC-UV-QTRAP6500+MS system sufficient to meet the updated analytical needs of
the parent project. The PI’s group has nearly two decades of strong expertise and hands-on operating
experience with LC-UV-MS/MS, thus will be able to quickly set up, maintain, and adjust the LC-UV-
QTRAP6500+MS system for the various methodologies and conditions optimal for the parent grant. In
summary, the purchase of the proposed QTRAP6500+ MS is vital for the ReLiM analyses proposed in the
parent grant. The pharmacokinetic data, including low but bioactive doses, acquired with the QTRAP6500+MS
provided by this grant will enable the PI’s group to develop the ReLiM-functionalized dressings, with no or
minimized graft-donor needs, capable of even better healing than was originally expected in the parent project.