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DNA-specific pattern recognition receptor activation following DNA electroporation

Award Number: R01CA196796
ORGANIZATION: NATIONAL CANCER INSTITUTE
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 02/01/2016
PERIOD OF PERFORMANCE END DATE: 01/31/2023

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DNA-specific pattern recognition receptor activation following DNA electroporation - DESCRIPTION (provided by applicant): Electroporation is a standard laboratory technique for transfection of bacteria, eukaryotic cells, and tissues in vivo. Our laboratories were among the first to develop in vivo plasmid electroporation for therapeutic applications by delivery to preclinical tumors, muscle, and skin and the first that conducted veterinary and clinical gene therapy trials. In vivo electroporation is becoming a well-accepted technique for clinical molecular delivery. Currently, nearly 100 clinical trials are registered in the NIH's clinicaltrials.gov database for electroporation of drugs and nucleic acids. We observed that complete tumor regression can occur when control backbone pDNA is electroporated into different tumors types using various electroporation protocols. Regression is preceded by the production of proinflammatory mRNAs and proteins. Our preliminary results implicate the activation of DNA-specific pattern recognition receptors (PRRs), which normally function for defense against pathogen invasion, in the induction of these proinflammatory proteins. These sensors are found in both immune and non-immune cell types. Activation of these sensors in non-immune cells, such as tumor cells, may contribute to the inflammation and regression that we observed in our previous research and may influence gene therapy efficacy. For example, inflammation may enhance the efficacy of an anti-tumor therapy or an infectious disease vaccine, but interfere with a protein replacement therapy targeting a healthy tissue. Our hypothesis is that DNA-specific PRR activation in non-immune cell types may influence the therapeutic potential of DNA electroporation. Therefore, in this proposal, we will elucidate the activation of PRRs in non-immune cells following different electroporation protocols in several normal cell types and in tumor cells, examine potential activation in a 3-dimensional microenvironment, and relate this pattern to subsequent in vivo effects. Our long-term goal is to be able to predict, based on putative PRR activation and in vivo effects, what electroporation parameters should be used for a specific tissue and therapeutic application. To test our hypothesis, we will conduct our experiments within the following specific aims: Specific Aim 1. Determine the DNA sensor activation pattern in response to plasmid DNA electroporation using various protocols in non-immune cell types Specific Aim 2. Determine effect of 3-dimensional culture and co-cultures of tumor and normal cells on the DNA sensor activation pattern in response to plasmid DNA electroporation Specific Aim 3. Determine the in vivo DNA sensor activation pattern after plasmid DNA electroporation and its potential influence on gene therapies delivered to tumor, muscle, and skin tissues.


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: 2024 ( Subtotal = -$13 )
2024	2020	UNIVERSITY OF SOUTH FLORIDA	4202 E FOWLER AVE	TAMPA	FL	33620	HILLSBOROUGH	USA	Cancer Treatment Research	000	5	11/21/2023	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	-$13
														Subtotal = -$13

Issue Date FY: 2022 ( Subtotal = $0 )
2022	2020	UNIVERSITY OF SOUTH FLORIDA	4202 E FOWLER AVE	TAMPA	FL	33620	HILLSBOROUGH	USA	Cancer Treatment Research	000	5	12/8/2021	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$0
														Subtotal = $0

Issue Date FY: 2020 ( Subtotal = $315,061 )
2020	2020	UNIVERSITY OF SOUTH FLORIDA	4202 E FOWLER AVE	TAMPA	FL	33620	HILLSBOROUGH	USA	Cancer Treatment Research	001	5	2/17/2020	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$315,061
2020	2020	UNIVERSITY OF SOUTH FLORIDA	4202 E FOWLER AVE	TAMPA	FL	33620	HILLSBOROUGH	USA	Cancer Treatment Research	004	5	5/4/2020	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$370,589
2020	2019	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	003	4	5/4/2020	NON-COMPETING CONTINUATION	-$306,683
2020	2019	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	000	4	2/17/2020	NON-COMPETING CONTINUATION	$0
2020	2018	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	002	3	5/4/2020	NON-COMPETING CONTINUATION	-$63,906
														Subtotal = $315,061

Issue Date FY: 2019 ( Subtotal = $306,683 )
2019	2019	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	000	4	12/20/2018	NON-COMPETING CONTINUATION	$306,683
														Subtotal = $306,683

Issue Date FY: 2018 ( Subtotal = $316,169 )
2018	2018	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	000	3	1/30/2018	NON-COMPETING CONTINUATION	$284,551
2018	2018	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	001	3	5/23/2018	NON-COMPETING CONTINUATION	$31,618
														Subtotal = $316,169

Issue Date FY: 2017 ( Subtotal = $316,169 )
2017	2017	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	001	2	6/7/2017	NON-COMPETING CONTINUATION	$31,618
2017	2017	OLD DOMINION UNIVERSITY	5115 HAMPTON BLVD	NORFOLK	VA	23529	NORFOLK CITY	USA	Cancer Treatment Research	000	2	1/20/2017	NON-COMPETING CONTINUATION	$284,551
														Subtotal = $316,169

Issue Date FY: 2016 ( Subtotal = $329,919 )
2016	2016	OLD DOMINION UNIVERSITY RESEARCH FOUNDATION	P.O. BOX 6369	NORFOLK	VA	23508	NORFOLK CITY	USA	Cancer Treatment Research	000	1	1/21/2016	NEW	$329,919
														Subtotal = $329,919

Grand Total All Awards = $1,583,988

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DNA-specific pattern recognition receptor activation following DNA electroporation

Award Number: R01CA196796

ORGANIZATION: NATIONAL CANCER INSTITUTE

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 02/01/2016

PERIOD OF PERFORMANCE END DATE: 01/31/2023

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