My Primary research interests involve (1) the molecular biology and genetics of cancer and (2) the isolation, culture, and characterization of adult tissue-derived stem cells, i.e., stem and multipotent progenitor cells other than embryonic stem cells. Current projects concern:

• Influence of genetic polymorphisms on Prostate Cancer risk and biology
• Pancreatic Islet isolation & culture; pancreatic epithelial cell culture and differentiation
• Isolation, improved culture, and multilineage differentiation of Adipose Stromal Cells, mesenchymal stem cells, and myoblasts
• Epithelial stem cell isolation & culture

1. Prostate Cancer Research
Epidermal growth factor (EGF) receptor intron 1 repeat polymorphisms in African-American and Caucasian males: Influence on prostate cancer risk or disease progression and interaction with androgen receptor CAG repeat polymorphisms:
This study involves the development of a prostate cancer database that will collect data not currently available on prostate cancer patients and their family members. Personal information and blood samples will be collected from all participants, and tissue samples will be collected from participants that undergo medically indicated biopsies or surgeries. I hypothesize that shorter CA repeats in intron 1 of the EGFR gene, by resulting in transcriptional enhancement of EGF receptor expression, will synergize with shorter CAG androgen receptor AR repeats to increase the risk of early onset prostate cancer and/ or promote the development of androgen-independent, aggressive prostate cancer. The status of Epidermal Growth Factor (EGF) Receptor intron 1 CA repeat and androgen receptor CAG repeat polymorphisms in African-American and Caucasian males will be determined by PCR analysis of DNA isolated from blood samples. These data will be studied in conjunction with the personal and medical information to determine whether the status of the EGF receptor polymorphism, alone or in combination with the androgen receptor polymorphism, influences the age of onset or biological characteristics (e.g., hormone dependence, invasiveness, metastasis) of prostate cancer.

2. Pancreatic Islet/Diabetes Research
Type 1 or Juvenile Diabetes is a serious disorder that usually appears at a young age. This form of diabetes occurs when the person's immune system attacks his or her own pancreas (autoimmune disease). Autoimmune destruction of insulin-producing beta cells in the pancreas (found in cell clusters called Islets of Langerhans) results in a lack of the hormone insulin, which regulates the uptake and utilization of blood sugar (glucose). This results in episodes of low blood sugar (hypoglycemia) and high blood sugar (hyperglycemia), which can be life threatening. Uncontrolled or poorly controlled diabetes can result in blindness, kidney disease, nerve damage that can lead to amputations, and increased risk of heart disease and stroke. Recently, islets isolated from pancreata of organ donors were infused into the liver of type I diabetes patients, and using a novel immuno-suppressive regimen, recipients attained insulin independence. While these results are quite promising, there will never be enough donor pancreata to treat even the most seriously ill type I diabetics. It is therefore vital to develop techniques that yield more islets from each donor pancreas, and to maintain their viability in culture before transplant. Even substantial improvements in these processes will not yield sufficient islets for all diabetics who could potentially benefit from this kind of procedure, so another important research area involves the cultivation of pancreatic ductal epithelial cells (or other stem cells) and their differentiation into islet cells in culture.
We therefore are focussing on the basic problems of improving islet viability in culture, and generating functional islet-like cell clusters from pancreatic precursors and stem cells in culture. Pancreatic-derived epithelial cells (PDECs) can be isolated from a "waste" fraction of islet preparations. A sub-population of these cells behaves as stem cells. That is, these cells can be expanded extensively in culture, and expanded PDECs can be induced to differentiate into beta-cell like cells, i.e. produce and secrete insulin. At Coriell, we have isolated islets and PDECs from pigs, and developed a novel, defined medium capable of sustaining the proliferation of the pancreatic epithelial cells from which islets develop. We have also obtained insulin-producing cells from these cultured stem cells. However, these processes are still inefficient, and much more work remains to be done in order to achieve results that would be clinically useful. Efforts to establish pancreatic stem cell cultures from non-human primates are currently underway, and this work will then inform our work with cells from human tissue.

3. Mesodermal Stem Cell Research
Dr. Moscatello's laboratory is also actively investigating "adult" tissue-derived Stem Cells from mesodermal tissues, i.e., (1) adipose stromal cells isolated from adipose tissue (fat); (2) mesenchymal stem cells isolated from bone marrow; (3) myoblasts, or muscle stem cells; and (4) umbilical cord blood stem cells. Stem cells are cells that are capable of long-term proliferation, and also of differentiation into multiple cell types. The two research areas that are key to the use of stem cells in clinical applications are (1) the ability to expand stem cell populations in culture, while still retaining their "stemness", or their ability to turn into multiple cell types, and (2) the conditions necessary to induce the cells to become the desired type of cell. Stem cells from several sources are currently being studied in the Differentiated Cell Laboratory. While some cells can only be obtained from tissues harvested after the death of a donor, other tissues can be obtained as biopsy or surgical waste. For instance, adipose tissue is waste from tumescent liposuction procedures; umbilical cord blood is obtained from the cord after the baby is born; and small amounts of muscle and bone marrow aspirate are obtained from diagnostic and surgical specimens. All these tissues contain stem cells capable of giving rise to their respective tissues of origin, and both adipose tissue and cord blood contain cells similar to the mesenchymal stem cells from bone marrow, which are capable of differentiation into adipocytes (fat), chondrocytes (cartilage), myocytes (muscle) and osteoblasts (bone). With the use of hormones, vitamins and growth factors, stem cells can become useful in treating particular diseases, whether caused by auto-immunity (such as diabetes), infectious disease, trauma, or the aging process. In order for these cells to achieve their clinical potential, our efforts focus on methods to optimize their isolation, culture, and cryopreservation using defined media.

4. Epidermal Stem Cell Research
Currently, almost all cultures in the Repositories are either lymphoblastoid cell lines or fibroblast lines and are prepared from peripheral blood samples and tissue biopsies, respectively. Both procedures are invasive, require professionals to acquire the sample and are moderately painful. In contrast, epidermal stem cells can be obtained from neonatal foreskins, obtained as waste from circumcisions, and potentially from hair follicles by plucking a few hairs! The locus of epidermal stem cells is the bulge that is part of the outer root sheath (ORS) (Rochat et al., 1994; Morris and Potten, 1994; reviewed in Alonso and Fuchs, 2003). These cells have extensive proliferative capacity, more than sufficient to establish, expand and distribute cell cultures.
The standard methods for culture of keratinocytes were developed by Rheinwald and Green (1975) using a feeder layer of irradiated fibroblasts. Refinements of these techniques have been developed to culture these cells to initiate cultures of human keratinocytes from the ORS which include the epidermal stem cells (Limat and Hunziker (1996)). This protocol takes advantage of the well-established ability of dermal fibroblasts to promote long-term expansion of epidermal precursor cells, which is necessary to obtain sufficient cells for distribution from the small inoculum that can be obtained from a few hair follicles. However, the presence of non-autologous fibroblasts could be a confounding factor for genetic analyses or clinical applications, so development of methods using defined media is an important focus of our research. Feeder layer-free culture protocols for epidermal stem cell culture have been developed including use of collagen coated plates and fibroblast-conditioned medium (Papini et al, 2003), or the addition of low concentrations of transforming growth factor beta 1 (TGFb1) (Fortunel et al (2003)). Until these recent developments, such cultures were restricted to growth for fewer than 20 population doublings, which would not be sufficient for the purposes of the Repository. These novel approaches may make it possible to culture epidermal precursor cells routinely as a third major and non-invasive method to the establishment of cultures for the Coriell Cell Repositories.

While pursuing graduate work at Purdue University, Dr. Moscatello ran a microbiology laboratory course as a full-time graduate instructor. After completing his doctoral studies in 1984, he continued teaching at Purdue. A few years later, he accepted a position as Assistant Professor of Microbiology at Richard Stockton College of New Jersey.

Following those experiences in teaching, he decided to re-enter research as a post-doctoral fellow at Thomas Jefferson University in Pennsylvania, where he worked in the laboratory of Dr. Albert Wong and was supported as an NIH post-doctoral research fellow studying the mechanisms by which a mutant epidermal growth factor receptor (EGFRvlll) results in the neoplastic transformation of cells. After several productive years, Dr. Moscatello was promoted to research instructor at the university where he continued his research into the signal transduction pathways activated downstream of EGFRvlll. However, considerable evidence strongly suggested that many aspects of signaling and transformation are quite different in epithelial and hematopoietic cells (cells that give rise to most human cancers) than in the fibroblastic cell lines commonly used in molecular biology studies. Attracted by the opportunity to work with such differentiated cells at the Coriell Institute, Dr. Moscatello accepted a position at the Institute in October 1999 to pursue his own research interests and to make such cell types more readily available to other biomedical researchers.

Dr. Moscatello is a member of the American Association for the Advancement of Science, the American Association for Cancer Research, and the American Society for Cell Biology.

Moscatello D.K., Dougherty M., Narins R.S., and Lawrence N.
Cryopreservation of human fat for soft tissue augmentation: Viability requires use of cryoprotectant and controlled freezing and storage. Derm. Surg. 31: 1506-1510, 2005.

Modjtahedi H., Moscatello D.K., Box G., Green M., Shotton C., Lamb D.J., Reynolds L.J., Wong A.J., Dean C., Thomas H., Eccles S.
Targeting of cells expressing wild-type EGFR and type-III mutant EGFR (EGFRvIII) by anti-EGFR MAb ICR62: A two-pronged attack for tumour therapy. Int. J. Cancer 105(2): 273-80, 2003.
PMID: 12673691 abstract

Antonyak M.A., Kenyon L.C., Godwin A.K., James D.C., Emlet D.R., Okamoto I., Tnani M., Holgado-Madruga M., Moscatello D.K., Wong A.J.
Elevated JNK activation contributes to the pathogenesis of human brain tumors. Oncogene 21(33): 5038-5046, 2002.
PMID: 12140754 abstract

Olapade-Olaopa, E.O., Ogunbiyi, J.O., MacKay, E.H., Muronda, C.A., Alonge, T.O., Danso, A.P., Moscatello, D.K., Sandhu, D.P., Shittu, O.B., Terry, T.R., Wong, A.J., and Habib, F.K.
Further characterization of storage-related alterations in immunoreactivity of archival tissue sections and its implications for collaborative multicenter immunohistochemical studies. Appl. Immunohistochem. Mol. Morphol. 3: 261-266, 2001.
PMID: 11556755 abstract

Moscatello, D.K. and Iozzo, R.V.
Interaction of Proteoglycans with Receptor Tyrosine Kinases. In Proteoglycan Protocols, Methods in Molecular Biology 171, Renato V. Iozzo, Ed. Humana Press, Totowa, New Jersey, pp. 427-434, 2001.

Tang, C.K., Gong, X-Q., Moscatello, D.K., Wong, A.J., and Lippman, M.E.
EGFRvIII enhances tumorigenicity in Human Breast Cancer. Cancer Research 60: 3081-3087, 2000.
PMID: 10850460 abstract

Olapade-Olaopa E.O., Moscatello, D.K., MacKay, E.H., Horsburgh, T., Sandhu, D.P.S., Terry, T.R., Wong A.J., and Habib F.K.
Evidence for the differential expression of a variant EGF receptor protein in human prostate cancer. British Journal of Cancer 82(1): 186-194, 2000.
PMID: 10638988 abstract

Maroun C.R., Moscatello, D.K., Naujokas, M.A., Holgado-Madruga, M., Wong, A.J., Park, M.
A conserved inositol phospholipid binding site within the pleckstrin homology domain of the Gab1 docking protein is required for epithelial morphogenesis. J. Biol. Chem. 274(44): 31719-31726, 1999.
PMID: 10531383 abstract  full text

Iozzo RV, Moscatello DK, McQuillan DJ, Eichstetter I
Decorin is a biological ligand for the epidermal growth factor receptor. J Biol Chem. 274(8):4489-92, 1999.
PMID: 9988678 abstract  full text

Antonyak MA, Moscatello DK, Wong AJ
Constitutive activation of c-Jun N-terminal kinase by a mutant epidermal growth factor receptor. J Biol Chem. 273(5):2817-22, 1998.
PMID: 9446590 abstract  full text

Moscatello DK, Santra M, Mann DM, McQuillan DJ, Wong AJ, Iozzo RV
Decorin suppresses tumor cell growth by activating the epidermal growth factor receptor. J Clin Invest. 101(2):406-12, 1998.
PMID: 9435313 abstract  full text

Moscatello DK, Holgado-Madruga M, Emlet DR, Montgomery RB, Wong AJ
Constitutive activation of phosphatidylinositol 3-kinase by a naturally occurring mutant epidermal growth factor receptor. J Biol Chem. 273(1):200-6, 1998.
PMID: 9417065 abstract  full text

Emlet, D.R., Moscatello, D.K., Ludlow, L.B., and Wong, A.J.
Subsets of epidermal growth factor receptors during activation and endocytosis. J. Biol. Chem. 272: 4079-4086, 1997.
PMID: 9020117 abstract  full text

Montgomery, R.B., Moscatello, D.K., Wong, A.J., and Stahl, W.L.
Epidermal growth factor receptor stimulation of diacylglycerol kinase. Biochem. Biophys. Res. Comm. 232: 111-116, 1997.
PMID: 9125112 abstract

Holgado-Madruga M, Moscatello DK, Emlet DR, Dieterich R, Wong AJ
Grb2-associated binder-1 mediates phosphatidylinositol 3-kinase activation and the promotion of cell survival by nerve growth factor. Proc Natl Acad Sci USA 94(23):12419-24, 1997.
PMID: 9356464 abstract  full text

Moscatello DK, Ramirez G, Wong AJ
A naturally occurring mutant human epidermal growth factor receptor as a target for peptide vaccine immunotherapy of tumors. Cancer Res. 57(8):1419-24, 1997.
PMID: 9108438 abstract

Moscatello, D.K., Montgomery, R.B., Sundareshan, P., McDanel, H., Wong, M.Y., and Wong, A.J.
Transformation and altered signal transduction by a naturally occurring mutant EGF receptor. Oncogene 13: 85-96, 1996.
PMID: 8700557 abstract

Holgado-Madruga, M., Emlet, D.R., Moscatello, D.K., Godwin, A.K., and Wong, A.J.
A Grb2-associated docking protein in EGF- and insulin-receptor signalling. Nature 379: 560-564, 1996.
PMID: 8596638 abstract

Montgomery, R.B., Moscatello, D.K., Wong, A.J., Cooper, J.A., and Stahl, W.L.
Differential modulation of MEK and MAP kinase activities by a mutant EGF receptor. J. Biol. Chem. 270: 30562-30566, 1995.
PMID: 8530489 abstract  full text

Moscatello, D.K., Holgado-Madruga, M., Godwin, A.K., Ramirez, G., Gunn, G., Zoltick, P.W., Biegel, J.A., Hayes, R.L., and Wong, A.J.
Frequent expression of a mutant Epidermal Growth Factor receptor in multiple human tumors. Cancer Res. 55: 5536-5539, 1995.
PMID: 7585629 abstract

Reist, C., Archer, G.E., Kurpad, S.N., Wikstrand, C.J., Vaidyanathan, G., Willingham, M.C., Moscatello, D.K., Wong, A.J., Bigner, D.D., and Zalutsky, M.R.
Tumor-specific anti-Epidermal Growth Factor receptor variant III monoclonal antibodies: Use of the tyramine-cellobiose radioiodination method enhances cellular retention and uptake in tumor xenografts. Cancer Res. 55: 4375-4382, 1995.
PMID: 7671250 abstract

Wikstrand, C.J., Hale, L.P., Batra, S.K., Hill, M.L., Humphrey, P.A., Kurpad, S.N., McLendon, R.E., Moscatello, D., Pegram, C.N., Reist, C.J., Traweek, S.T., Wong, A.J., Zalutsky, M.R., and Bigner, D.D.
Monoclonal antibodies against EGFRvIII are tumor specific and react with breast and lung carcinomas and malignant gliomas. Cancer Res. 55: 3140-3148, 1995.
PMID: 7606735 abstract