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RECOGNITION OF COMMENSAL AND PATHOGENIC BACTERIA BY ORAL EPITHELIUM

Award Number: R01DE018307
ORGANIZATION: NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 07/01/2007
PERIOD OF PERFORMANCE END DATE: 06/30/2012

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RECOGNITION OF COMMENSAL AND PATHOGENIC BACTERIA BY ORAL EPITHELIUM - DESCRIPTION (provided by applicant): The goal of this proposal is to address at the molecular and cellular level the differential recognition of commensal and pathogenic bacteria by gingival epithelial cells. The oral epithelium is in direct contact with bacteria and plays an important role in the innate immune response by the expression of mediators and antimicrobial factors. Paradoxically, it is typically in a non-inflamed state despite constant exposure to bacteria. Consistent with this observation, Preliminary Data demonstrate that a pathogenic bacterium, P. gingivalis generally upregulates cytokine expression in oral epithelial cell cultures in contrast to a commensal bacteria, ¿ gordonii which does not. Mucosal epithelium in the gut is also able to maintain homeostasis while exposed to a high commensal bacterial load, yet retains the capacity to respond to pathogenic bacteria by upregulating an inflammatory response. The proposed studies will use liquid-air interface cultures of primary human gingival epithelial cells which form multi-layers of highly differentiated cells. Three representative bacteria will be examined: a well defined commensal, S. gordonii, an opportunistic commensal, F. nucleatum and a pathogenic bacteria, P. gingivalis. mRNA profiling will build upon Preliminary Data to establish whether this spectrum of bacteria exhibit significant differences in upregulating the innate immune response focusing on antibacterial factors including defensins, cytokines/chemokines and their receptors, pattern recognition receptors, intracellluar signalling molecules, and molecules that affect barrier function. Results will be verified for selected genes by real-time PCR and at the protein level. Aim 2 will establish whether each bacterium induces different signalling pathways in the epithelial culture system described for Aim 1. These studies will examine MAP kinase signalling and activation of transcription factors. Their functional importance will be assessed by use of siRNA or specific inhibitors. The second aspect of Aim 2 is to identify differential induction of signalling complexes by identifying molecules that associate with TLR2 on bacterial stimulation. The approach will use immunoaffinity minicolumns and mass-spectroscopy. Once identified, siRNA approaches will be used to investigate their functional significance. Aim 3 will utilize gnotobiotic mice to test the hypothesis that acquisition of a commensal bacteria at infancy attenuates the later response to pathogenic bacteria. The goal of this Aim is to investigate whether age-associated acquisition of a commensal flora induces bacterial tolerance.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2011 ( Subtotal = $295,665 )
2011	2011	UNIVERSITY OF PENNSYLVANIA	3451 WALNUT STREET	PHILADELPHIA	PA	19104	PHILADELPHIA	USA	Oral Diseases and Disorders Research	000	6	6/15/2011	NON-COMPETING CONTINUATION	$295,665
														Subtotal = $295,665

Issue Date FY: 2010 ( Subtotal = $304,810 )
2010	2010	UNIVERSITY OF PENNSYLVANIA	3451 WALNUT STREET	PHILADELPHIA	PA	19104	PHILADELPHIA	USA	Oral Diseases and Disorders Research	000	5	9/7/2010	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$304,810
2010	2009	UNIVERSITY OF MEDICINE & DENTISTRY OF NEW JERSEY	65 BERGEN STREET	NEWARK	NJ	07101	ESSEX	USA	Oral Diseases and Disorders Research	001	4	9/7/2010	NON-COMPETING CONTINUATION	$0
														Subtotal = $304,810

Issue Date FY: 2009 ( Subtotal = $292,847 )
2009	2009	UNIVERSITY OF MEDICINE & DENTISTRY OF NEW JERSEY	65 BERGEN STREET	NEWARK	NJ	07107	ESSEX	USA	Oral Diseases and Disorders Research	002	4	7/20/2009	NON-COMPETING CONTINUATION	$302,389
2009	2008	UNIVERSITY OF MEDICINE & DENTISTRY OF NEW JERSEY	65 BERGEN STREET	NEWARK	NJ	07107	ESSEX	USA	Oral Diseases and Disorders Research	001	3	3/16/2009	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$302,388
2009	2008	BOSTON UNIVERSITY	25 BUICK ST	BOSTON	MA	02215	SUFFOLK	USA	Oral Diseases and Disorders Research	000	2	3/16/2009	NON-COMPETING CONTINUATION	-$311,930
														Subtotal = $292,847

Issue Date FY: 2008 ( Subtotal = $311,931 )
2008	2008	BOSTON UNIVERSITY	25 BUICK STREET	BOSTON	MA	02215	SUFFOLK	USA	Oral Diseases and Disorders Research	000	2	6/13/2008	NON-COMPETING CONTINUATION	$311,931
														Subtotal = $311,931

Issue Date FY: 2007 ( Subtotal = $331,025 )
2007	2007	BOSTON UNIVERSITY	25 BUICK STREET	BOSTON	MA	02215	SUFFOLK	USA	Oral Diseases and Disorders Research	000	1	6/28/2007	NEW	$331,025
														Subtotal = $331,025

Grand Total All Awards = $1,536,278

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RECOGNITION OF COMMENSAL AND PATHOGENIC BACTERIA BY ORAL EPITHELIUM

Award Number: R01DE018307

ORGANIZATION: NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 07/01/2007

PERIOD OF PERFORMANCE END DATE: 06/30/2012

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