David Kirsch
Principal Investigator
Barbara Levine University Distinguished Professor
Professor of Radiation Oncology
Professor of Pharmacology and Cancer Biology
Member of the Duke Cancer Institute
Affiliate of the Regeneration Next Initiative

Kirsch Lab Overview

David Kirsch and Yvonne Mowery in the Kirsch Lab

The Kirsch Lab uses sophisticated genetically engineered mouse models, cellular and molecular biology and biochemistry to study cancer and radiation biology. Our research ranges from fundamental basic questions into the mechanisms by which tumor suppressor genes prevent cancer to translational projects that focus on mechanisms of metastasis, tumor response to radiation therapy and normal tissue injury from radiation. In a collaborative environment, we work together to make discoveries and strive to translate our research into clinical trials for patients with cancer.

David Kirsch, MD, PhD, is the Barbara Levine University Professor at Duke. As a physician-scientist, he cares for bone and soft tissue sarcoma patients with radiation therapy in the Duke Cancer Institute. His research program spans the continuum from basic cancer research, to translational radiation biology, to clinical trials, such as SU2C-SARC032 that is testing the combination of immunotherapy and radiation therapy for high risk extremity sarcoma. His lab research is supported by an R35 Outstanding Investigator Award from the National Cancer Institute, grants from the National Institute of Allergy and Infectious Disease and the Department of Defense, as well as several philanthropic organizations including StandUp2Cancer, the Gilbert Family Foundation, the Slifka Foundation and the Leon Levine Foundation. Follow Dr. Kirsch on Twitter here

Research

Genetically Engineered Mouse Models (GEMMs) to Study Cancer and Radiation Biology

Our laboratory uses GEMMs to model cancer because the tumors develop in their native microenvironment in animals with an intact immune system to closely resemble the natural history of human tumors. These advantages over traditional transplanted tumor models make GEMMs ideal preclinical platforms to dissect mechanisms of tumor biology and develop novel treatments. We have pioneered the development of spatially and temporally restricted mouse models of cancer using Cre-LoxP technology, dual recombinase technology, and CRISPR/Cas9 genome editing. We have used these models to study metastasis, to develop intraoperative molecular imaging, and to dissect the mechanisms of tumor and stromal cells to radiation therapy. We have also used GEMMs to reveal mechanisms by which the p53 tumor suppressor protein regulates acute toxicity and late effects of radiation therapy in specific cell types.

GEMMs cancer and radiation biology

 

Publications

Torok JA, Oh P, Castle KD, Reinsvold M, Ma Y, Luo L, Lee CL, Kirsch DG . Deletion of ATM in tumor but not endothelial cells improves radiation response in a primary mouse model of lung adenocarcinoma. Cancer Research. 2018 Oct 12. doi: 10.1158/0008-5472.CAN-17-3103. PMID:30315114

Huang J, Chen M, Whitley MJ, Kuo HC, Xu ES, Walens A, Mowery YM, Van Mater D, Eward WC, Cardona DM, Luo L, Ma Y, Lopez OM, Nelson CE, Robinson-Hamm JN, Reddy A, Dave SS, Gersbach CA, Dodd RD, Kirsch DG. Generation and comparison of CRISPR-Cas9 and Cre-mediated genetically engineered mouse models of sarcoma. Nature Communications. 2017 Jul 10;8:15999. PMCID:PMC5508130

Whitley MJ, Cardona DM, Spasojevic I, Ferrer JM, Cahill J, Lee CL, Snuderl M, Blazer DG 3rd, Hwang SE, Greenup RA, Mosca PJ, Mito JK, Cuneo KC, Larrier NA, O’Reilly EK, Riedel RF, Eward WC, Strasfeld DB, Fukumura D, Jain RK, Lee WD, Griffith LG, Bawendi MG, Kirsch DG, Brigman BE. A Mouse-Human Phase I Co-Clinical Trial of the Protease-Activatable Fluorescent Probe LUM015 for Intraoperative Imaging of Cancer. Science Translational Medicine. 2016 Jan 6; 8(320): 320ra4.  PMCID:PMC4794335

Moding EJ, Castle KD, Perez BA, Oh P, Min HD, Norris H, Ma Y, Cardona DM, Lee CL, Kirsch DG. Tumor Cells, but not Endothelial Cells, Mediate Eradication of Primary Sarcomas by Stereotactic Body Radiation Therapy. Science Translational Medicine.  2015 Mar 11;7(278): 278ra34. PMCID:PMC4360135

Moding EJ, Lee CL, Castle KD, Oh P, Mao L, Zha S, Min HD, Ma Y, Das S, Kirsch DG. Atm deletion with dual recombinase technology preferentially radiosensitizes tumor endothelium. Journal of Clinical Investigation. 2014 Aug;124(8):3325-38. PMCID:PMC4109553

Lee CL, Moding EJ, Cuneo KC, Li Y, Sullivan JM, Mao L, Washington I, Jeffords LB, Rodrigues RC, Ma Y, Das S, Kontos CD, Kim Y, Rockman HA, Kirsch DG.  p53 Functions in Endothelial Cells to Prevent Radiation-Induced Myocardial Injury in Mice. Science Signaling. 2012 Jul 24; 5(234). PMCID:PMC3533440

Kirsch DG, Santiago PM, di Tomaso E, Sullivan JM, Hou WS, Dayton T, Jeffords LB, Sodha P, Mercer KL, Cohen R, Takeuchi O, Korsmeyer SJ, Bronson RT, Kim CF, Haigis KM, Jain RK, Jacks T.  p53 Controls Radiation-Induced Gastrointestinal Syndrome in Mice Independent of Apoptosis. Science. 2010 Jan 29; 327(5965):593-6. PMCID:PMC2897160

Kirsch DG, Dinulescu DM, Miller JB, Grimm J, Santiago PM, Young NP, Nielsen GP, Quade BJ,  Chaber CJ, Schultz CP, Takeuchi O, Bronson RT, Crowley D, Korsmeyer SJ, Yoon SS, Hornicek FJ, Weissleder R, Jacks, T.  A Spatially and Temporally Restricted Mouse Model of Soft Tissue Sarcoma.  Nature Medicine. 2007;13:992-997.*=equal contribution. PMID:17676052

 


Current Research Projects in the Kirsch Lab

 

Mechanisms of Response and Resistance to Radiation and Immune Checkpoint Blockade

Since the initial clinical trials of immune checkpoint blockade in patients with melanoma, immunotherapy, such as anti-programmed cell death protein 1 (PD-1), has been approved for many cancers and demonstrated impressive responses in some patients. However, the majority of cancer patients fail to respond to immune checkpoint blockade alone. In preclinical studies of mouse tumors transplanted into syngeneic mice, numerous investigators have reported dramatic responses to immune checkpoint blockade combined with radiation therapy, but these tumor models do not develop in a native microenvironment under immunosurveillance. Although genetically engineered mouse models (GEMMs) of cancer do co-evolve with a native immune system, these tumors have a very low number of somatic mutations. Therefore, they express limited neoantigens and generally do not engage effector T cells.

Mice with conditional mutations

To overcome the limitations of both transplant and GEMM models for studying immunotherapy, we have generated a novel autochthonous mouse model of high mutational load sarcoma initiated by deletion of p53 and a chemical carcinogen.  Using this novel highly mutated autochthonous tumor model, we are investigating mechanisms by which primary tumors respond or are resistant to immunotherapy and radiation therapy.  We are also testing whether this treatment combination not only improves local control of the irradiated tumor, but also the subsequent development of lung metastasis. This work has the potential to lead to new approaches to activate the immune system and prevent the development of metastasis.
 

Defining a Genetic Signature for Radiation-Induced Cancer

Radiation-induced sarcomas are a significant clinical problem because treatment of the normal tissue with a second course of radiation therapy increases the risk of a serious radiation-related toxicity. It is difficult to determine whether a second cancer that arises in a patient after radiation therapy is a new primary tumor, a recurrence of the original tumor, or a radiation-induced cancer. Because of this uncertainty, clinically the term “radiation-associated” is utilized rather than “radiation-induced."

To define a genetic signature of radiation-induced cancer, we are performing whole exome sequencing of mouse sarcomas that develop after focal, high dose radiation therapy. We are comparing the genetic signature to our other mouse models of sarcoma induced by Cre recombinase and/or carcinogens.  We are also seeking to identify gene mutations in sarcomas that arise specifically after radiation therapy.

Sarcoma cohort and mouse genotype with tumor initiation

 

Sensitizing Brainstem Glioma to Radiation Therapy

Diffuse intrinsic pontine glioma (DIPG), also referred to as high-grade brainstem glioma, is a pediatric cancer that accounts for the majority of deaths from brain tumors in children. Radiotherapy is the standard of care for DIPG, as the anatomic location of the tumor precludes surgery and no chemotherapeutic agents have shown efficacy over radiation alone. Despite routine treatment with radiotherapy, fewer than 10% of patients survive two years from diagnosis. Using established GEMMs of brainstem glioma with different tumor suppressor genes, we are applying the Cre-loxP system to delete kinases in the tumor cells that we hypothesize will radiosensitize brainstem gliomas and improve survival after radiotherapy.

Diagram of genetically engineered mice

 

Other Research Projects

  • Mechanisms by which the tumor suppressor p53 prevents cancer
  • Mechanisms by which the tumor suppressor ATRX prevents cancer
  • Mechanisms by which FUS-CHOP drives the development of cancer and radiosensitivity
  • Mechanisms of metastasis
  • Mechanisms of normal tissue injury after radiation
  • Cancer metabolism and radiation response
  • Epigenetic regulators of tumorigenesis

Publications

Bookshelf with books

Van Mater D, Xu E, Reddy A, Añó L, Sachdeva M, Huang W, Williams N, Ma Y, Love C, Happ L, Dave S, Kirsch DG.  Injury promotes sarcoma development in a genetically and temporally restricted manner.  JCI Insight. 2018 Oct 18;3(20) doi: 10.1172/jci.insight.123687. [Epub ahead of print]. PMID:30333301

Torok JA, Oh P, Castle KD, Reinsvold M, Ma Y, Luo L, Lee CL, Kirsch DG.  Deletion of ATM in tumor but not endothelial cells improves radiation response in a primary mouse model of lung adenocarcinoma. Cancer Research. 2018 Oct 12. doi: 10.1158/0008-5472.CAN-17-3103. PMID:30315114

Brownstein JM, Wisdom AJ, Castle KD, Mowery YM, Guida P, Lee CL, Tommasino F, Tessa CL, Scifoni E, Gao J, Luo L, Campos LDS, Ma Y, Williams N, Jung SH, Durante M, Kirsch DG. Characterizing the potency and impact of carbon ion therapy in a primary mouse model of soft tissue sarcoma. Molecular Cancer Therapuetics. 2018 Apr;17(4):858-868. PMCID:PMC5912881

Huang J, Chen M, Whitley MJ, Kuo HC, Xu ES, Walens A, Mowery YM, Van Mater D, Eward WC, Cardona DM, Luo L, Ma Y, Lopez OM, Nelson CE, Robinson-Hamm JN, Reddy A, Dave SS, Gersbach CA, Dodd RD, Kirsch DG. Generation and comparison of CRISPR-Cas9 and Cre-mediated genetically engineered mouse models of sarcoma. Nature Communications. 2017 Jul 10;8:15999. PMCID:PMC5508130

Dodd RD, Lee CL, Overton T, Huang W, Eward WC, Luo L, Ma Y, Ingram DR, Torres KE, Cardona D, Lazar AJ, Kirsch DG. NF1+/- Hematopoietic cells accelerate malignant peripheral nerve sheath tumor development without altering chemotherapy response. Cancer Research. 2017 Aug 15;77(16):4486-4497. PMCID:PMC5839126

Sachdeva M, Mito JK, Lee CL, Zhang M, Li Z, Dodd RD, Cason D, Luo L, Ma Y, Van Mater D, Gladdy R, Lev DC, Cardona DM, Kirsch DG. MicroRNA-182 drives metastasis of primary sarcomas by targeting multiple genes. Journal of Clinical Investigation. 2014 Oct 1; 124(10):4305-4319. PMCID:PMC4191039

Whitley MJ, Cardona DM, Spasojevic I, Ferrer JM, Cahill J, Lee CL, Snuderl M, Blazer DG 3rd, Hwang SE, Greenup RA, Mosca PJ, Mito JK, Cuneo KC, Larrier NA, O’Reilly EK, Riedel RF, Eward WC, Strasfeld DB, Fukumura D, Jain RK, Lee WD, Griffith LG, Bawendi MG, Kirsch DG*^, Brigman BE*.A mouse-human phase 1 co-clinical trial of a protease-activated fluorescent probe for imaging cancer. Science Translational Medicine. 2016 Jan 6; 8(320): 320ra4.  PMCID:PMC4794335 *co-senior authors, ^corresponding author

Lee CL, Castle KD, Moding EJ, Blum JM, Williams N, Luo L, Ma Y, Borst L, Kim Y, Kirsch DG. Acute DNA damage activates the tumor suppressor p53 to promote radiation-induced lymphoma. Nature Communications. 2015 Sep 24;6:8477. PMCID:PMC4586051

Dodd RD, Sachdeva M, Mito JK, Eward WC, Brigman BE, Ma Y, Dodd L, Kim Y, Lev D, Kirsch DG. Myogenic transcription factors regulate pro-metastatic miR-182. Oncogene. 2016 Apr 7;35(14):1868-1875.PMCID:PMC4523886

Moding EJ, Castle KD, Perez BA, Oh P, Min HD, Norris H, Ma Y, Cardona DM, Lee CL, Kirsch DG. Tumor Cells, but not endothelial cells, mediate eradication of primary sarcomas by stereotactic body radiation therapy. Science Translational Medicine.  2015 Mar 11;7(278): 278ra34. PMCID:PMC4360135

Van Mater D, Añó L, Blum JM, Webster MT, Huang W, Williams N, Ma Y, Cardona DM, Fan CM, Kirsch DG. Acute tissue injury activates satellite cells and promotes sarcoma formation via the HGF/c-MET signaling pathway. Cancer Research. 2015 Feb 1;75(3):605-14. PMCID:PMC4327867

Sachdeva M, Mito JK, Lee CL, Zhang M, Li Z, Dodd RD, Cason D, Luo L, Ma Y, Van Mater D, Gladdy R, Lev DC, Cardona DM, Kirsch DG. MicroRNA-182 drives metastasis of primary sarcomas by targeting multiple genes. Journal of Clinical Investigation. 2014 Oct 1; 124(10):4305-4319. PMCID:PMC4191039

Moding EJ, Lee CL, Castle KD, Oh P, Mao L, Zha S, Min HD, Ma Y, Das S, Kirsch DG. ATM deletion with dual recombinase technology preferentially radiosensitizes tumor endothelium. Journal of Clinical Investigation. 2014 Aug 1; 124(8):3325-38. PMCID:PMC4109553

Lee CL, Moding EJ, Kirsch DG. Reining in radiation injury: HIF2α in the gut. Science Translational Medicine. 2014 May 14;6(236):236fs20.doi:10.1126/scitranslmed.3009155. PMID:24828075

Blum JM, Añó L, Li Z, Van Mater D, Bennett BD, Sachdeva M, Lagutina I, Zhang M, Mito JK, Dodd LG,  Cardona DM, Dodd RD, Williams N, Ma Y, Lepper C,  Linardic CM, Mukherjee S, Grosveld GC, Fan CM, Kirsch DG. Distinct and overlapping sarcoma subtypes initiated from muscle stem and progenitor cells. Cell Reports. 2013 Nov 27; 5(4):933-940. PMCID: PMC3893104

Lee CL, Moding EJ, Cuneo KC, Li Y, Sullivan JM, Mao L, Washington I, Jeffords LB, Rodrigues RC, Ma Y, Das S, Kontos CD, Kim Y, Rockman HA, Kirsch DG.  p53 functions in endothelial cells to prevent radiation-induced myocardial injury in mice. Science Signaling. 2012 Jul 24; 5(234). PMCID:PMC3533440

​For a complete list of publications, please click here. 

People

Mentored Junior Faculty

Butler Harris Assistant Professor in Radiation Oncology
Assistant Professor of Radiation Oncology

Lab Members

Program Coordinator
Research Scientist
Postdoctoral Associate
Postdoctoral Associate
Research Associate Senior
Resident, Radiation Oncology, Class of 2024
Graduate Student
Graduate Student
Graduate Student
Graduate Student
Graduate Student
Undergraduate Student
Research Technician
Research Technician
Research Technician
Research Technician
Research Technician

 

Kirsch Lab group photo

Protocols

p53 FRT Genotyping

 

Schematic of the allele

p53 FRT Genotyping

 

Primers

#1: 5’-CAAGAGAACTGTGCCTAAGAG-3’
#2: 5’-CTTTCTAACAGCAAAGGCAAGC-3’
#3: 5’-ACTCGTGGAACAGAAACAGGCAGA-3’

 

Reaction Components

Reagent Volume (ul)
10x buffer 2
10 mM dNTP mix 0.4
20 uM F Primer 0.4
20 uM R Primer 0.4
Taq DNA Polymerase 1
Nuclease free dH2O 11.8
Genomic DNA (~200 ng) 4
Total 20


Cycling
p53FRT (Primers #1 and #2) 

Step Temp (C)
1 94
2 94
3 55
4 72
Back to step 2, 38 times  
5 72
6 4

 

Expected Results

p53FRT

Mutant = 292 bp
Heterozygote = 292 and 258 bp
Wild Type = 258 bp

 

p53Δ2-6 (Primers #1 and #3)

Step Temp (C)
1 94
2 94
3 55
4 72
Back to step 2, 38 times  
5 72
6 4

 

Expected Results

p53Δ2-6:
Recombined allele = 352 bp
Unrecombined or WT = No band

Mouse codon-optimized Flp (FlpO) is described in the following reference:
Raymond, C.S., and Soriano, P. (2007). High-efficiency FLP and PhiC31 site-specific recombination in mammalian cells. PLoS One 2, e162.

Events

Lab Outing 2019 – Urban Axes, Durham, NC

Lab Outing 2019 – Urban Axes, Durham, NC

 

Lab Outing 2018 — Xtreme Park Adventures, Durham, NC

Lab Outing 2018 – Xtreme Park Adventures, Durham, NC

 

Lab Outing 2017 – U.S. National Whitewater Center, Charlotte, NC​

Lab Outing 2017 – U.S. National Whitewater Center, Charlotte, NC​

 

Lab Outing 2016 – Great Escape, Durham, NC

Lab Outing 2016 – Great Escape, Durham, NC

 

Lab Outing 2015 – ZipQuest, Raleigh, NC

Lab Outing 2015 – ZipQuest, Raleigh, NC

 

Lab Outing 2014 — Scavenger Hunt, North Carolina Museum of Science, Raleigh, NC

Lab Outing 2014 – Scavenger Hunt, North Carolina Museum of Science, Raleigh, NC

 

Lab Outing 2013 — Frankie's Fun Park, Raleigh, NC

Lab Outing 2013 – Frankie's Fun Park, Raleigh, NC

 

Lab Outing 2012 — U.S.National Whitewater Center, Charlotte, NC

Lab Outing 2012 – U.S.National Whitewater Center, Charlotte, NC

Contact

Mailing Address

Kirsch Lab
Duke University Medical Center
DUMC Box 91006
Durham, NC 27708

Shipping Address

450 Research Drive
LSRC Bldg, Room B324
Durham, NC 27708