Rice, Robert - Research

Areas of Interest

1. Proteomic profiling of hair, nail, epidermis. For the past 15 years, my laboratory has been heavily engaged in keratinocyte proteomics. We have exploited mass spectrometry to analyze the previously unknown isopeptide cross-linked proteins in structures composed of corneocytes. We presented the first cross-linked proteome of hair shaft and followed that with analyses of nail plate and epidermal callus. We have shown this approach can be applied to humans and to animal models of disease (mouse epidermis, nail, hair). This approach, useful to understand the downstream phenotypic consequences of damage even to a single gene, is projected to permit more quantitative monitoring of skin disease after therapeutic treatment. We have explored applications to characterizing effects of environmental exposures (arsenic, TCDD) that can be modeled in cell cultures and to analysis of genetically variant peptides, assisting individual identification in forensic investigation.

  • Lee YJ, Rice RH, Lee YM (2006) Proteome analysis of human hair shaft: From protein identification to posttranslational modification. Molec Cell Proteomics 5:789-800
  • Rice RH, Xia Y, Alvarado RJ, Phinney BS (2010) Proteomic analysis of human nail plate. J Proteome Res 9:6752-6758
  • Rice RH, Bradshaw KM, Durbin-Johnson BP, Rocke DM, Eigenheer RA, Phinney BS, Sundberg JP (2012) Differentiating inbred mouse strains from each other and those with single gene mutations using hair proteomics. PLoS One 7:12 e51956
  • Rice RH, Durbin-Johnson BP, Mann SM, Salemi M, Urayama S, Rocke DM, Phinney BS, Sundberg JP (2018) Corneocyte proteomics: Applications to skin biology and dermatology. Exp Dermatol 27(8):931-938

2. Mechanism of arsenic carcinogenicity. The mystery of how inorganic arsenic contributes to skin cancer is gradually being solved. Reports that arsenite can reduce the effectiveness of DNA repair (PARP-1) provide a plausible but only a partial explanation. Our work traces important signaling pathways with which arsenite interferes, thereby preserving the germinative capability of the cells. This includes markedly prolonging EGF receptor activity, preventing its suppression by insulin/IGF action. Arsenite also suppresses Notch-1 activation, protein kinase Cδ expression and dual specificity phosphatase induction, all of which inhibit cells from leaving the mitotic pool and thus inhibit differentiation. We also showed that arsenite [As(III)] ultimately is the active oxidation state, but arsenate [As(V)] approaches its potency upon intracellular reduction to arsenite. These findings help rationalize the contribution of inorganic arsenic in the water supply to cancer development. We have now found antimonite [Sb(III)] mimics arsenite remarkably, suggesting it also targets the epidermis.

  • Reznikova TV, Phillips MA, Patterson TJ, Rice RH (2010) Opposing actions of insulin and arsenite converge on PKCδ to alter keratinocyte proliferative potential and differentiation. Molec Carcinogenesis 49:398-409
  • Phillips MA, Qin Q, Hu Q, Zhao B, Rice RH (2013) Arsenite suppression of BMP signaling in human keratinocytes. Toxicol Appl Pharmacol 269:290-296
  • Phillips MA, Cánovas A, Wu P-W, Islas-Trejo A, Medrano JF, Rice RH (2016) Parallel responses of human epidermal keratinocytes to inorganic SbIII and AsIII. Environ Chem 13:963-970
  • Phillips MA, Cánovas A, Rea MA, Islas-Trejo A, Medrano JF, Durbin-Johnson B, Rocke DM, Rice RH (2020) Deducing signaling pathways from parallel actions of arsenite and antimonite in human epidermal keratinocytes. Sci Rep 10:2890

3. Development of keratinocyte culture models to study toxic exposures. We initially found that epithelial cells from a variety of rat tissues could be serially cultured and routinely developed into continuous lines, serving as a culture model for squamous metaplasia. We also derived a spontaneously immortalized keratinocyte line from ostensibly normal human epidermis and have used it as a minimally-deviated complement to normal human epidermal cells for studies of differentiation and response to carcinogens such as arsenic, benzo(a)pyrene, 2,3,7,8-tetrachlorodibenzo-p-dioxin and smoldering woodsmoke.

  • Phillips MA, Rice RH (1983) Convergent differentiation in cultured rat cells from nonkeratinized epithelia:  Keratinocyte character and intrinsic differences. J Cell Biol 97:686-691
  • Rice RH, Steinmann KE, deGraffenried LA, Qin Q, Taylor N, Schlegel R (1993) Elevation of cell cycle control proteins during spontaneous immortalization of human keratinocytes.  Molec Biol Cell 4:185-194
  • Hu Q, Rice RH, Qin Q, Phinney BS, Eigenheer RA, Bao W, Zhao B (2013) Proteomic analysis of human keratinocyte response to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure. J Proteome Res 12:5340-5347
  • Chan LK, Nguyen KQ, Karim N, Yang Y, Rice RH, He G, Denison MS, Nguyen TB (2019) Relationship between the molecular composition, visible light absorption, and health-related properties of smoldering woodsmoke aerosols. Atmos Chem Phys 20:539-559

4. Role of keratinocyte transglutaminase (TGM1). We showed that cross-linked envelopes in mature corneocytes consists of isopeptide cross-linked protein containing a previously unknown protein, involucrin. We also characterized and molecularly cloned the TGM1 transglutaminase enzyme responsible for the cross-linking. We provided the monoclonal antibody we raised to TGM1 to help identify defects in this enzyme as the most common cause of lamellar ichthyosis. Our cDNA for the full length coding region was instrumental in demonstrating that its expression in ichthyosis keratinocytes could repair the defect. We have shown that TGM1 in teleosts is membrane-bound, suggesting it was re-purposed during evolution to help adapt to the terrestrial environment.

  • Thacher SM, Rice RH (1985) Keratinocyte-specific transglutaminase of cultured human epidermal cells: Relation to cross-linked envelope formation and terminal differentiation. Cell 40:685-695
  • Phillips MA, Stewart BE, Rice RH (1992) Genomic structure of keratinocyte transglutaminase. Recruitment of new exon for modified function. J Biol Chem 267:2282-2286
  • Phillips MA, Qin Q, Mehrpouyan M, Rice RH (1993) Keratinocyte transglutaminase membrane anchorage: Analysis of site-directed mutants. Biochemistry 32:11057-11063
  • Rodriguez Cruz SI, Phillips MA, Kültz D, Rice RH (2017) Tgm1-like transglutaminases in Tilapia (Oreochromis mossambicus). PLoS One 12(5):e0177016

5. Applications of proteomics to personal identification using hair and epidermal trace evidence. Intrigued by our finding that inbred mouse strains could be distinguished by their pelage hair proteomes, we found that individual humans (each analogous to an inbred mouse strain) could be distinguished as well. Although this protein profiling appeared promising for application to forensic science, we recognized that an even better approach (with random match probabilities now in the millions) is provided by use of genetically variant peptides in hair shaft digests reflecting SNPs in the donor genome. In collaboration with Dr. Glendon Parker, we have developed this methodology and shown its applicability not only to the hair shaft but also to fingerprints and other bodily tissues and fluids.

  • Parker GJ, Leppert T, Anex DS, Hilmer JK, Matsunami N, Baird L, Stevens J, Parsawar K, Durbin-Johnson BP, Rocke DM, Nelson C, Fairbanks DJ, Wilson AW, Rice RH, Woodward SR, Bothner B, Hart B, Leppert M (2016) Demonstration of protein-based human identification using the hair shaft proteome. PLoS One 11(9):e0160653
  • Wu P-W, Mason KE, Durbin-Johnson BP, Salemi M, Phinney BS, Rocke DM, Parker GJ, Rice RH (2017) Proteomic analysis of hair shafts from monozygotic twins: Expression profiles and genetically variant peptides. Proteomics 17, 13–14, 2017, 1600462
  • Borja T, Karim N, Goecker Z, Salemi M, Phinney BS, Naeem M, Rice RH, Parker GJ (2019) Proteomic genotyping of fingermark donors with genetically variant peptides. For Sci Intl: Genet 42:21-30
  • Plott TJ, Karim N, Durbin-Johnson BP, Swift DP, Youngquist RS, Salemi M, Phinney BS, Rocke DM, Davis MG, Parker GJ, Rice RH (2020) Age-related changes in hair shaft protein profiling and genetically variant peptides. For Sci Int Gen 47:102309

6. Proteomic manifestations of genetic defects. During the past 8 years, my lab has engaged in collaborations to discern alterations in proteomic profiles in genetic disease or genomic alteration of epidermis and adnexa. Efforts with 10 different groups have resulted in 14 publications. Financial support has included contributions from consortia such as FIRST (Foundation for Ichthyosis and Related Skin Types) and PC (Pachyonychia Congenita) Project.

  • Rice RH, Durbin-Johnson BP, Ishitsuka YI, Salemi M, Phinney BS, Rocke DM, Roop DR (2016) Proteomic analysis of loricrin knockout mouse epidermis. J Proteome Res 15:2560-2566
  • Rice RH, Durbin-Johnson BP, Salemi M, Schwartz ME, Rocke DM, Phinney BS (2017) Proteomic profiling of pachyonychia congenita plantar callus. J Proteomics 165:132–137
  •      Jaeger K, Sukseree S, Zhong S, Phinney BS, Mlitz M, Buchberger M, Narzt MS, Gruber F, Tschachler E, Rice RH, Eckhart L (2019) Cornification of nail keratinocytes requires autophagy for bulk degradation of intracellular proteins while sparing components of the cytoskeleton. Apoptosis 24:62-73
  •      Karim N, Durbin-Johnson B, Rocke DM, Salemi M, Phinney BS, Naeem M, Rice RH (2019) Proteomic manifestations of genetic defects in autosomal recessive congenital ichthyosis. J Proteomics 201:104-109

Complete List of Published Work in PubMed (145 entries): https://www.ncbi.nlm.nih.gov/pubmed/?term=rice+rh