F32 Trial and Error: Lessons Learned from the Application Process Kimberly Brothers Ph.D. Research Instructor Shanks lab University of Pittsburgh Department of Ophthalmology Charles T. Campbell Laboratory

Outline Letters of Support Aims Research Strategy Goals for fellowship and career Activities planned under this award Reviewer comments Introduction to resubmission

Your PI wants you to write a F32 or better, YOU want to write one What do I do first? Try to get a copy of someone’s F32 Download SF424 instructions https://grants.nih.gov/grants/guide/pa-files/PA-16-307.html (look for SF424 instructions)

Letters of Support Ask people who have trained you for letters of support Give them plenty of time to submit the letters

Biosketch

Biosketch Contributions to Science

MARKETING PORTFOLIO BE HONEST: What are my strengths? What are my weaknesses? Do I have meritorious publications that demonstrate my ingenuity/talent? Are there cool techniques proposed for the project that I would benefit learning? Do I have sufficient preliminary data to support the aims and goal(s) of the project?

You Must Focus!!! Subject [# Awards] Dry Eye [90] Dry Eye and Infection [22] Dry Eye and Bacteria [2]

I Found My Niche/Topic Now What? Training Environment Postulating testable hypothesis Training Plan

Specific Aims Should be independent Not very time intensive Facilities and equipment that will support aims IACUC/IRB if there is animal/human work

Aims and Research Strategy Aim 1. Identify the bacterial secreted factor(s) responsible for reduced corneal cell migration. Aim 2. Test the hypothesis that bacteria inhibit corneal wound healing in vivo. Good to have 3 aims because you can always cut one.

Research Strategy Aim 1: Identify the bacterial secreted factor(s) responsible for reduced corneal cell migration SM transposon mutagenesis (1a) Biochemical approach HPLC-MS, NMR (1b) Protein/Compounds screen (1c) Pitfalls: Use transcriptomic approach RNA- Seq

Research Strategy Aim 2: Test the hypothesis that bacteria inhibit corneal wound healing in vivo

Aims and Research Strategy Aim 1. Identify the bacterial secreted factor(s) responsible for reduced corneal cell migration. Aim 2. Test the hypothesis that bacteria inhibit corneal wound healing in vivo. Aims not independent

Updated Aims Aim 1. Test the hypothesis that bacteria inhibit corneal wound healing in vivo Aim 2. Test the hypothesis that S. marcescens (SM) strain PIC3611 lipopolysaccharide (LPS) inhibits corneal wound healing

Revised Research Strategy Test the hypothesis that bacteria inhibit corneal wound healing in vivo (Aim 1) Test the hypothesis that S. marcescens (SM) strain PIC3611 lipopolysaccharide (LPS) inhibits corneal wound healing (Aim 2) Complementation of LPS mutants (Aim 2.1) Test non-viable bacterial ability to inhibit wound healing (Aim 2.2) Biochemical approach (deplete secretomes of LPS, purify LPS) (Aim 2.3)

Revised Research Strategy Preliminary Data

Goals for Fellowship and Career Simplified into research goals and professional development goals http://www.regionplus.co.uk

Goals for Fellowship and Career Simplified into research goals and professional development goals Simplified into research goals and professional development goals http://www.regionplus.co.uk

Goals for Fellowship and Career Simplified into research goals and professional development goals Simplified into research goals and professional development goals http://www.regionplus.co.uk

Activities Planned Under this Award

Activities Planned Under this Award Not so many classes and incorporate milestones for career advancement http://p.vitalmx.com

Sponsor Training Plan Lists all past trainees Environment, Research Facilities Plan for trainee Meet every Monday regarding project Biology of Vision course Dept Seminars Presenting at local and national meetings Publishing manuscripts

Sponsor Training Plan If PI is junior faculty or topic outside lab focus add co-sponsor List other grants/Funds

Responsible Conduct of Research

Vertebrate Animals

Vertebrate Animals

Equipment

Facilities

You Don’t Want an Angry Review Committee

You Want One That is Receptive to Your Idea(s)

Choosing your study section https://public.csr.nih.gov/StudySections/Fellowship/Pages/default.aspx

Reviewer comments PI has not trained enough PhD’s (add a co-sponsor) Too ambitious Concerned about funding status of sponsor and co- sponsor (letter from Research Chair) Wanted annual milestones to ensure proper training

Introduction to Resubmission

Cover Letter

Cover Letter

Cover Letter