INSERT TITLE HERE

MY NAME
MY AFFILIATION

2018/10/23

An NIH R01 proposal, submitted in response to

PAR-XX-XXX: INSERT TITLE OF PAR HERE.

Compiled on 2018/10/23 at 07:12:00.

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INSERT DATE HERE

Dear NIH:

I am submitting a grant proposal entitled “INSERT TITLE HERE.”

• Please assign this application to the National Institute for FIXME

• Please assign this application to the following study section: FIXME

I have discussed this application with INSERT NAME OF PROGRAM OFFICER at INSERT NAME OF INSTI-
TUTE. They recommended the above two assignments.
Thank you very much.

Sincerely,

INSERT NAME
INSERT AFFILIATION
Project Summary/Abstract
Project narrative
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Specific Aims
Hyperoxia-induced oxidative stress contributes to the disordered lung development that leads to bronchopul-
monary dysplasia (BPD) in prematurely born extremely low birth weight (ELBW) infants.1 We have recently
observed endothelial mitochondrial DNA (mtDNA) dam-age and bioenergetic dysfunction in infants who develop
BPD.2 In our novel Mitochondrial-Nuclear eXchange (MNX) murine BPD model, mice with C57 mtDNA [C57
wildtype (WT) (C57 (n)uclear DNA /C57 (mt) DNA) and C3HMNX mice (C3Hn/C57mt)] had increased hyper-
oxic lung injury when compared to mice with C3H mtDNA [C3HWT (C3Hn/C3Hmt) and C57MNX (C57n/C3Hmt)]
(manuscript in review).3 These findings indicate that mtDNA damage and polymorphisms are associated with
BPD pathogenesis, as has been noted in other pulmonary oxidative disor-ders. However, mechanisms through
which mtDNA damage and inherited mtDNA polymor-phisms (haplotypes) mediate BPD are unknown. MtDNA
is sensitive to oxidative mutations and requires frequent repair by the DNA glycosylase OGG1.4 MtDNA-damage
associated mo-lecular patterns (DAMP) from endothelial cells and activated platelets accumulate in lungs and
cause inflammatory damage in pneumonia and sepsis.58 Ethnically variant mtDNA hap-lotypes such as H (Euro-
pean) and L (African) cause differential electron transport chain (ETC) complex bioenergetic function by modifying
nuclear (possibly through epigenetic changes) and mitochondrial genetic expression of these proteins.9, 10 There
is a critical need to explore such mtDNA-mediated mechanisms in neonatal oxidative lung injury because lack
of such information could limit development of mitochondrial-specific therapies for BPD. Our long-term goal is to
harness mitochondrial function to minimize BPD severity. Our cur-rent objectives in pursuit of this goal are to (i)
identify mechanisms through which mtDNA damage and mtDNA polymorphisms modify neonatal oxidative lung
injury and (ii) test the utility of mtDNA as a BPD biomarker. Our central hypothesis is that oxidant stress causes
mtDNA-damage mediated inflammation and mtDNA haplogroup-mediated ETC dysfunction to amplify neonatal
lung injury (Figure 1). The rationale for this project is that identifying such mtDNA-mediated mechanisms can
provide the basis for evaluating novel strategies such as DNase therapy and personalized risk stratification based
on mtDNA haplotype in infants at risk for BPD. We will achieve our objectives through the following Specific Aims:
1. Evaluate mtDNA damage as a neonatal oxidative lung injury amplifier. OGG1 KO mice with deficient mtDNA
repair, mice with deficient platelet activation and controls will be exposed to air or hyperoxia. We hypothesize
increased pulmonary cell-free mtDNA content and damage in OGG1 KO mice and decreased circulating mtDNA-
DAMP content in mice with deficient platelet activation vs. their controls along with corresponding changes in
lung injury and inflammation.
2. Identify mechanisms of mtDNA haplotype-induced neonatal oxidative lung injury. C57WT, C3HWT, C57MNX
and C3HMNX mice will be exposed to air or hyperoxia. We hy-pothesize decreased C-IV or C-I activity in hyper-
oxic mice with C57 vs. C3H mtDNA and cor-responding lung injury, gene expression and methylation changes.
3. Define biomarkers for BPD risk based on platelet mtDNA and activation. We will evaluate a prospective
cohort of ELBW infants for platelet activation as well as for plasma mtDNA content. We hypothesize that these
measures will be increased in BPD-susceptible infants vs. infants who survive without BPD.
We anticipate significant advancement of the field through our expected outcomes of dis-covering a) mech-
anisms through which mtDNA modifies pathogenesis of neonatal oxidative lung injury and b) validity of plasma
mtDNA as a BPD biomarker. Innovations include use of novel transgenic models to study BPD. As principal inves-
tigator I have performed and pub-lished studies on several experiments outlined in our approach. The research
environment will include mentors who are experts in pulmonary and mitochondrial biology. This AHA award will
improve my expertise in these areas and enable me to successfully compete for NIH funding.
Research strategy
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Specific Aim 1:

Specific Aim 2:
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