David M. Kaetzel, PhD

Pedigo, N., Zhang, X.-H., Bruno, E.C., Kaetzel, C.S., Dugan, A., Koszewski, N. and Kaetzel, D.M.  A 5’-distal enhanceosome in the PDGF-A gene is activated in choriocarcinoma cells via ligand-independent binding of vitamin D receptor and constitutive jun kinase signaling.  Oncogene 24:2654-2666, 2005.  PMID: 15829977 

Ma, D., Nutt, C.L., Shanehsaz, P., Peng, X., Louis, D. and Kaetzel, D.M.   Autocrine PDGF-dependent gene expression in glioblastoma cells is mediated largely by activation of the transcription factor SRE-BP, and is associated with altered genotype and patient survival in human brain tumors.  Cancer Research 65: 5523-5534, 2005.  PMID: 15994924 (no PMCID to date)

Kaetzel, D.M., Zhang, Q., Yang, M., McCorkle, J.R., Craven, R.  Potential roles of 3’-5’ exonuclease activity of NM23-H1 in DNA repair and malignant progression. J. Bioenerg. Biomembr. 38: 163-167, 2006. PMID: 17039395 (no PMCID to date)

Kowluru, A., Veluthakal, R. and Kaetzel, D.M.  Regulatory roles for nm23/nucleoside diphosphate kinase-like enzymes in insulin secretion from the islet β cell. J. Bioenerg. Biomembr. 38: 227-232, 2006. PMID: 16957985 (no PMCID to date)

Pedigo, N., Zhang, H., Mishra, A., McCorkle, R., Ormerod, A.K. and Kaetzel, D.M.   Retinoic acid inducibility of the human PDGF-A gene is mediated by 5'-distal DNA motifs that overlap with basal enhancer and vitamin D response elements.  Gene Expr. 14: 1-12, 2007.  PMID: 17933214

Kaetzel, D.M., Xing, E.X., Pedigo, N.G. and Ormerod, K.  The calcitriol analogue EB1089 impairs alveolarization and induces localized regions of increased fibroblast density in neonatal rat lung.    Exp. Lung. Res. 34: 155-182, 2008.  PMCID: PMC2747600

Mishra, A., Spear, B.T., Kraner, S.D. and Kaetzel, D.M. PDGF-A promoter and enhancer elements provide efficient and selective antineoplastic gene therapy in multiple cancer types.  Cancer Gene Ther. 16:298-309, 2009.  PMCID: PMC2730454

Srinivasan, D., Kaetzel, D.M. and Plattner, R.  Reciprocal regulation of Abl and receptor tyrosine kinases.  Cell Signal. 21:1143-1150, 2009.  PMCID: PMC2701649

Kaetzel, D.M., McCorkle, J.R., Novak, M., Yang, M. and Jarrett, S.  Potential contributions of antimutator activity to metastasis suppressor function of NM23-H1.  Mol. Cell. Biochem. 329:161-165, 2009. PMCID: PMC2747477

Novak, M., Jarrett, S.G., McCorkle, J.R., Mellon, I. and Kaetzel, D.M.  Multiple mechanisms underlie metastasis suppressor function of NM23-H1 in melanoma.  Naunyn-Schmied. Arch. Pharmacol. 384:433-438, 2011.  PMID: 21448569 (no PMCID to date).

Jarrett, S.G., Novak, M., Harris, N., Mellon, I., Zhang, Q., Arnaud-Dabernat, S., Daniel, J.-Y., Ciesielski, M.J., Fenstermaker, R.A. and Kaetzel, D.M.  The metastasis suppressor NM23-H1 promotes repair of UV-induced DNA damage and suppresses UV-induced melanomagenesis. Cancer Res. 72: 133-143, 2012.  PMCID: PMC3251703

Ganguly, S., Fiore, L.S., Sims, J.T., Friend, J.W., Srinivasan, D., Cibull, M.L., Wang, C., Novak, M., Kaetzel, D.M. and Plattner, R. Activation of c-Abl and Arg in human melanoma cells promotes survival, proliferation, invasion, and metastasis via distinct molecular pathways. Oncogene 31:1804-1816, 2012.  PMCID: PMC3235241

Deng, X., Li, Q., Hoff, J., Yang, H., Jin, H., Erfani, S.F., Sharma, C., Zhou, P., Rabinovitz, I., Sonnenberg, A., Yi, A., Zhou, P., Stipp, C.S., Kaetzel, D.M., Hemler, M.E. and Yang, X.H.  Integrin-associated CD151 drives ErbB2-evoked mammary tumor onset and metastasis.  Neoplasia 14:678-689, 2012.  PMCID: PMC3431176

Veluthakal, R., Kaetzel, D. and Kowluru, A. NM23-H1 regulates glucose-stimulated insulin secretion in pancreatic beta cells via Arf6-Rac1 signaling axis.  Cell. Physiol. Biochem., 32:533-541, 2013. PMCID: PMC3845215

Fiore, L.S., Ganguly, S.S., Sledziona, J., Cibull, M.L., Wang, C., Richards, D.L., Neltner, J.M., Beach, C., McCorkle, J.R.,  Kaetzel, D.M. and Plattner, R. c-Abl and Arg induce cathepsin-mediated lysosomal degradation of the NM23-H1 metastasis suppressor in invasive cancer.  Oncogene 33:4508-4520, 2014.   PMCID: PMC3979510

Hoff, J.T., Baldwin, L.A., Lefringhouse, J., Zhang, M., Liu, Z., Erfani, S., She, Q.-B., Jia, C., Ueland, F.R., van Nagell, J.R., Wang, C., Xu, M., Kaetzel, D.M., Liu, C., Luo, J., Drapkin, R., Zhou, B. and Yang, X.H. CD151 regulates ovarian tumor growth and progression. Oncotarget 5:12203-12217, 2014. PMCID: PMC4322965

Kaetzel, D.M., Leonard, M.K., Cook, G.S., Novak, M., Jarrett, S.G., Yang, X. and Belkin, A.M. Dual functions of NME1 in suppression of cell motility and enhancement of genomic stability in melanoma.  Naunyn-Schmied. Arch. Pharmacol. 388:199-206, 2015. PMCID: PMC4294989

Progression of cancer to lethal forms requires the acquisition of driver mutations and aberrant programs of gene expression that endow cells with the ability to grow at distant sites, a process known as metastasis.  Current cancer therapies often fail as they are almost exclusively directed to the hyperproliferative phenotype of primary tumor cells, and do not effectively target the metastatic process.  Clearly, there is a dire need for new treatment options to target cancer in its advanced forms.  The primary thrust of my National Cancer Institute-funded research program has been the elucidation of molecular mechanisms that mediate cancer progression and metastasis, and has centered to a large extent on the NME/NM23 family of metastasis suppressor genes.  Our studies of the NME1 isoform has yielded considerable insights into molecular mechanisms that suppress metastatic potential, with one of the most significant being our identification of a novel mechanism underlying metastasis-driving mutations in melanoma. 

The NME Family of Metastasis Suppressor Genes 

Metastasis suppressors inhibit the metastatic activity of cancer cells with little effect on primary tumor growth  (Smith and Theodorescu, 2009). Mouse NME1 was the first metastasis suppressor gene to be described, with confirmation of suppressor activity of its human homolog nm23-h1 in cell lines and in melanoma, breast carcinoma and other cancers (Hartsough and Steeg, 2000). Although mechanisms underlying metastasis suppressor activity of the NM23-H1/M1 protein are not well understood, it harbors three distinct enzymatic activities that might mediate its antimetastatic functions. First to be described was its nucleoside diphosphate kinase (NDPK) activity, which maintains balance in nucleotide pools by catalyzing transfer of -phosphate between NDPs and NTPs (Agarwal et al., 1978). NM23-H1 also possesses a histidine kinase activity that may mediate an anti-motility function of the molecule (Wagner et al., 1997).

Our laboratory was the first to describe an association of 3’-5’ exonuclease activity with the NME1 protein (Ma et al., 2004), and we subsequently showed this activity to be essential for its metastasis suppressor function (Zhang et al., 2011). 3’-5’ exonucleases provide proofreading during DNA replication and repair (Shevelev and Hübscher, 2002), and consistent with this function, we showed the NM23 homolog in S. cerevisiae possesses antimutator activity (Yang et al., 2009). More recently, we have also demonstrated antimutator activity of NME1 in UVR-treated melanoma cell lines, and that NME1 specifically promotes both the nucleotide excision/NER (Jarrett et al., 2011) and double-strand break/DSBR (unpublished observations) pathways for DNA repair. Importantly, NME1 deficiency is associated with increased rates of spontaneous mutations in a variety of melanoma and nontransformed cell lines, strongly suggesting its expression may be essential for suppression of progression- and metastasis-driving mutations. Importantly, we have recently demonstrated a direct physical association of NME1 with sites of DNA damage, indicating its direct participation in the DNA repair process.  Using a transgenic mouse strain harboring a concurrent deficiency in both the NME1 or NME2 genes, we observe vulnerability to ultraviolet light-induced melanoma in situ, consistent with a DNA repair function in vivo (Jarrett et al., 2011).  Moreover, this NME1-deficient genotype confers aggressive metastasis when bred into a mouse model of melanoma with otherwise low metastatic potential (Jarrett et al., 2013). Taken together, our work has revealed an unexpected and significant mechanistic role for NME1 proteins in both initiation and progression of melanoma.

Current Projects 

In a project funded by the National Cancer Institute (R01 CA159871, “Suppression of Melanoma Initiation and Progression by NM23-H1”), we have proposed three aims to better understand how NME deficiency drives progression of melanoma to metastatic forms.  The first aim will employ transgenic mice harboring ablations of either the NME1 or NME2 loci, or tandem ablation of both loci, to measure their individual and combined contributions to metastasis suppression.  The second aim will probe more deeply into the molecular mechanisms through which NME proteins participate in the NER and DSBR pathways of DNA repair, including assessment of the individual roles played by the NDPK and 3’-5’ exonuclease functions.  We have recently demonsrtated for the first that NME1 is recruited to sites of DNA damage, where it interacts physically with other proteins (e.g. XRCC4, RAD50) involved in DSBR. Developing and applying NME1 mutants that disrupt various aspects of its repair function, we plan to determine the contribution of the DNA repair function of NME1 to metastasis suppressor activity in melanoma cell lines and transgenic mice.  In perhaps the most ambitious and exciting aim, we are applying the powerful technologies of whole genome sequencing and RNA-seq to identify candidate metastasis-driving genomic alterations and gene expression profiles in the melanoma lesions of our transgenic mouse model.  These studies are being conducted in collaboration with Dr. John Carpten at TGen (Phoenix), and preliminary results have already identified a number of unique mutations in these metastatic melanoma tumors that represent excellent candidates for further study.  For example, we have identified an indel in the Grin2a locus, which encodes an NMDA receptor isoform recently identified by exome sequencing to be a potential metastasis driver in human melanoma (Wei et al., 2011).  The detection of the Grin2a mutation in metastatic melanomas of our NM23 knockout mice indicates the relevance of this model to the human disease, and its immediate promise for identifying other novel metastasis-driving events caused by NM23 deficiency.

Our laboratory is also highly focused on understanding the molecular mechanisms underlying the anti-invasive activity of NM23 proteins.  Microarray analysis of our NM23-H1-transfected melanoma cell lines has revealed its potent regulation of a broad spectrum of mRNAs, with siRNA and forced expression approaches validating the functional relevance of interesting candidate mRNAs, encoding such molecules as LRP1b, aldolase C and integrin beta 3.  In addition, we have determined that NM23-H1 upregulates expression of fibronectin, which we have found to be essential for motility-suppressing interactions with a4b1 integrin (Novak et al., in preparation).  These observations have opened a wealth of opportunities to better understand how NM23 acutely suppresses the metastatic process. We have also identified a critical role of NME1 as a survival factor for melanoma stem cells and are actively engaged in efforts to examine the implications of this finding for regulating metastatic potential of melanoma cells.

We are also actively engaged in a project to exploit recominant NME1 as a culture medium supplement for expansion of human mesenchymal stem cells. A key aim to to expand the cells while maintaining their pluripotency, with the long-term goal of providing MSCs for regenerative therapies in human disase states. This line of Research is funded by the Maryland Stem Cell Research Fund through 2018. 

Awards and Affiliations

1978                Hubbard Scholarship Award, University of Maryland

1983-1985       NRSA Postdoctoral Fellowship (AM06981), Case Western Reserve University

1998-present   American Association for Cancer Research

2000                C. T. Wethington, Jr. Award for Research Excellence, College of Medicine, University of Kentucky.

2004-present   Metastasis Research Society

2011-present   Melanoma Research Society

2014-present   Pan-American Society for Pigment Cell Research

2007-2011      Standing Member, NIH Peer Review, Cancer Genetics Study Section (CG)

2009               Full Member, NIH Peer Review, Topics in CG Study Section ARRA (ZRG1 OBT-P (95) S)

2011               Full Member, NIH Peer Review, Topics in Cancer Genetics Study Section (ZRG1 CG-A (02) S); Full Member NIH Peer Review, SEP/SRG ZRG1 OBT-Z (02) M

2013               DOD Peer Review, Breast Cancer Research Program, IDX-PBY Panel (panel title confidential)

2013-2016      DOD Peer Review, IdeaA-SF/CDA Panel (panel title confidential)

2015, 2016      NIH Peer Review, Tumor Progression and Metastasis Study Section (TPM), ad hoc

2015                Chair, DOD Peer Review, IdeaA-SF/CDA Panel (panel title confidential)

9/10/12- 6/30/17 (P.I.)
"Suppression of Melanoma Initiation and Progression by NM23-H1"
National Cancer Institute, 1 R01 CA159871-01A1
Annual Direct Costs (year 1): $364,248 Total Costs: $1,483,233

7/1/15- 6/30/17          

Graduate Mentor (Minority Student Supplement)

“Suppression of Melanoma Initiation and Progression by NM23-H1”

National Cancer Institute, 3R01CA159871-04S1                             

Annual Direct Costs (year 1):  $34,061

7/1/2015- 6/30/2018    (P.I.)

“Enhanced Expansion of Undifferentiated, Pluripotent Human Mesenchymal Stem Cells by NME Proteins”

Maryland Stem Cell Foundation (MSCRF), 2015-MSCRFI-1638

Annual Direct Costs (year 1):  $200,000

Total Direct Costs: $600,000

State-of-the-art molecular biological techniques (e.g. plasmid construction, measurement of steady-state levels for nucleic acids and proteins) Technologies for forced expression and siRNA-mediated silencing of gene expression Identifying protein-protein interactions (e.g. pull-downs, yeast 2-hybrid) Chromatin immunoprecipitation Static and live microscopic imaging (e.g. confocal, TIRF) for assessing cell motility NextGen sequencing (whole genome, RNAseq) High-throughput screening for metastasis-inhibiting compounds HPLC for protein purification (state-of-the-art Waters system) Mouse models for metastatic melanoma, including transgenic mouse strains for spontaneous melanoma and explantation of cell lines in immunodeficient and syngeneic strains Methodology for measuring activity of DNA repair pathways (NER, DSBR, BER, etc.)