GRANT WRITING

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GRANT WRITING
Specific Aims
Tim Weaver
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Parts of the Grant

• Abstract
• Specific Aims
• Background and Significance
• Preliminary Studies/Progress Report
• Research Design

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The Writing Order

• Specific Aims
• Research Design
• Preliminary Studies/Progress Report
• Background and Significance
• Abstract

Plan the experiments to determine if the specific
aim is feasible
What preliminary data do I need to support the
feasibility of each specific aim?
What background do I need to understand the
specific aims?
Always write the abstract last
Always begin with the specific aims
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In the beginning.

• A critical idea is the single most important
  element of the grant application
• The idea must be
• Original
• Nontrivial
• Add significant new knowledge or fill an existing
  knowledge gap

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Specific Aims

• This page is a road map of your grant
• First impressions are important
• Write your aims again and again and again

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Structure of a specific aim

• Short introductory paragraph
• Brief overview of project
• Significance
• Central hypothesis or goal
• Specific aims
• Descriptive one line title
• (key preliminary data supporting hypothesis)
• Hypothesis/goal
• Experimental approach (how you will test the
  hypothesis)
• Summary sentence (why this experiment is
  important)

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Specific Aim Introductory paragraph

• Although SP-C is the first surfactant component
  to appear during lung development, the function
  of SP-C is the least understood of the surfactant
  proteins. Several lines of evidence strongly
  support the hypothesis that SP-C is important for
  surfactant function and turnover Replacement
  surfactants with recombinant SP-C as the sole
  protein component have excellent surface
  properties in vitro and in vivo SP-C facilitates
  the uptake of lipids by Type II cells and, most
  importantly, several term human infants with
  normal levels of SP-A and SP-B but undetectable
  SP-C protein rapidly developed severe RDS and
  ultimately succumbed to the disease. The overall
  goal of this project is to assess the role of
  SP-C in surfactant homeostasis (see Fig.1). SP-C
  function(s) will be identified by systematically
  altering the structure of the molecule and
  evaluating changes in surfactant assembly,
  secretion, biophysical activity, recycling and
  catabolism. The central hypothesis of this
  proposal is that the extramembrane domains of
  SP-C are important for intracellular trafficking,
  packaging of lamellar body phospholipids and/or
  secretion whereas the transmembrane domain of
  SP-C is essential for the biophysical properties
  of the peptide and/or uptake of alveolar
  phospholipids.

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Structure of a specific aim

• Specific Aim 1. Identification of Motif(s)
  Required for Intracellular Trafficking of SP-C.
  This aim will test the hypothesis that the
  sorting determinant(s) which directs SP-C to the
  regulated secretory pathway is located at the
  NH2-terminal end of the mature peptide. Deletion
  constructs of SP-C will be cloned in frame with
  green fluorescent protein (GFP) and transiently
  transfected into PC-12 cells. Intracellular
  trafficking of SP-C GFP will be monitored by
  confocal microscopy. These studies will identify
  the peptide domain(s) which is essential for
  trafficking of SP-C to dense core granules of the
  regulated secretory pathway.

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Structure of a specific aim

• Specific Aim 2. Identification of Motif(s)
  Required for Secretion of SP-C. SP-C is a
  transmembrane protein which is secreted into the
  airway. This aim will test the hypothesis that
  inward vesiculation of the SP-C proprotein in
  multivesicular bodies is essential for secretion
  of SP-C and that the NH2- and/or COOH-terminal
  propeptide is required for inward vesiculation.
  Deletion constructs of the SP-C proprotein will
  be transiently transfected into isolated Type II
  epithelial cells and the kinetics of SP-C
  secretion assessed by immunoprecipitation.
  Inward vesiculation will be assessed by
  immunogold labeling of ultrathin cryosections.
  These studies will provide insight into the
  mechanism whereby an integral membrane protein
  can be secreted.

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Structure of a specific aim

• Specific Aim 3. The Effect of Palmitoylation on
  SP-C Trafficking and Surfactant Homeostasis.
  This aim will test the hypothesis that
  palmitoylation of Cys5-Cys6 and/or Lys11 is
  important for packaging of surfactant
  phospholipids in lamellar bodies we will also
  test the hypothesis that palmitoylation alters
  SP-C mediated uptake of surfactant phospholipids
  from the alveolus. The three amino acids in SP-C
  known to be palmitoylated will be individually or
  combinatorially mutated to prevent acylation.
  Peptide trafficking, lipid packaging, and
  secretion of non-palmitoylated SP-C will be
  evaluated in transiently transfected Type II
  epithelial cells. In order to assess the
  importance of SP-C palmitoylation for surfactant
  homeostasis, palmitoylation mutants will be
  expressed in SP-C knockout mice. The
  pathophysiology associated with the loss of SP-C
  palmitoylation will provide new insight into the
  physiologic importance of this post-translational
  modification for surfactant homeostasis.

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Structure of a specific aim

• Specific Aim 4. The Role of the Transmembrane
  Domain in Surfactant Homeostasis. This aim will
  test the hypothesis that the amino acid
  composition, and in particular the valine
  content, of the membrane spanning domain is
  critical to the function of SP-C. SP-C
  constructs in which the transmembrane has been
  selectively mutated will be expressed in SP-C
  knockout mice. Mutations resulting in
  perturbation of surfactant homeostasis will
  provide new insight into the function of SP-C and
  the structural basis for this function.

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Hypothesis formulation

• It is not enough to formulate the hypothesis- you
  must rigorously test it
• Design experiments that establish cause-effect
• Experiments that establish a correlatioon do not
  rigorously test the hypothesis

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Remember.

• Hypotheses are frequently wrong
• Structure your experiment so that a negative
  outcome is informative
• Propose an alternative hypothesis
• Specific Aim 1
• If the sorting motif is not in the N-terminal
  peptide then we will test the hypothesis that the
  C-terminal peptide is critical for sorting of
  SP-C to the secretory granule

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Example hypotheses

• We will test the hypothesis that EGF treatment is
  correlated with SP-C expression
• We will test the hypothesis that expression of
  SP-C is developmentally and spatially regulated
• We will test the hypothesis that annexin I
  mediates lamellar body formation and surfactant
  recycling in alveolar type II cells and the
  annexin I-mediated processes are under growth
  factor and cellular calcium control

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Potential problem areas

• How many specific aims
• Integration of specific aims
• Contingent aims
• Relationship of specific aims to central
  hypothesis
• Goals vs hypotheses
• Depth vs breadth
• Novelty
• Lack of hypotheses
• Alternative hypotheses
• Organization
• Physiological/biological/clinical relevance

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A Word to the Wise.

• Read and follow the instructions
• Misspellings, grammatical errors and incorrect
  references reflect badly on your judgment
• Grant writing is the process of educating the
  reviewer
• NEVER assume the reviewer knows what you mean