This proposal aims to characterize novel basement membrane zone molecules associated with anchoring filaments and to determine the supramolecular structure and intermolecular interactions of anchoring filament components with lamina dense and hemidesmosomal components. The fundamental hypothesis of this project is that by elelucidating the structure and function of anchoring filaments we will be able to determine the molecular basis of junctional epidermolysis bullosa (JEB) and discover new genes which are affected in this disorder. Preliminary data are resented on three novel anchoring filaments proteins, LAD-1 (mAb 123 antigen), LH39 antigen and laminin 6, which are the three best candidates for new gene mutations in JEB. LAD-1 is a 120 kD upper anchoring filament protein which appears to be involved in insertion of anchoring filaments to the hemidesmosome and is absent in a subset of patients with generalized benign atrophic JEB (GABJEB). LH39 antigen is a lower anchoring filament protein which appears to be involved in insertion of anchoring filaments to the lamina densa and which is absent is a subset of patients with HBEB. Laminin-6 forms a disulfide complex with laminin-5 anchoring filaments, contains a novel a chain and could contain the primary defect in a subset of JEB patients. We propose to determine the entire cDNA sequence of the LH39 antigen and to propose additional experiments to determine its function and potential involvement in JEB. We similarly propose methods to determine the structure of the laminin 6 a chain. The interactions of anchoring filament components with hemidesmosome and lamina densa components will be analyzed by several methods, including solid state, chromatogrphic and centrifugation based ligand binding assays, cell binding assays and an in vitro basement membrane assembly model. Additionally, we propose to characterize the phenotypic features of GABJEB keratinocytes, to tranfect LAD-1 and BP180cDNA to effect phenotypic reversion in these cells and in conjunction with project 3, to study the effects of site directed mutagenesis in JEB keratinocytes using LAD-1, BP180 and laminin-5 cDNA. At the end of the proposed funding period, we hope to have significantly elucidated the basis of dermal-epidermal cohesion across the lamina lucida and to have demonstrated significant new mechanisms involving new candidate genes in JEB, setting the stage for further progress in gene therapy.

Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Stanford University
United States
Zip Code
Vijayakumar, Soundarapandian; Dang, Suparna; Marinkovich, M Peter et al. (2014) Aberrant expression of laminin-332 promotes cell proliferation and cyst growth in ARPKD. Am J Physiol Renal Physiol 306:F640-54
Gao, Jing; DeRouen, Mindy C; Chen, Chih-Hsin et al. (2008) Laminin-511 is an epithelial message promoting dermal papilla development and function during early hair morphogenesis. Genes Dev 22:2111-24
Tran, Mark; Rousselle, Patricia; Nokelainen, Pasi et al. (2008) Targeting a tumor-specific laminin domain critical for human carcinogenesis. Cancer Res 68:2885-94
Waterman, Elizabeth A; Sakai, Noriyasu; Nguyen, Ngon T et al. (2007) A laminin-collagen complex drives human epidermal carcinogenesis through phosphoinositol-3-kinase activation. Cancer Res 67:4264-70
Pullar, Christine E; Baier, Brian S; Kariya, Yoshinobu et al. (2006) beta4 integrin and epidermal growth factor coordinately regulate electric field-mediated directional migration via Rac1. Mol Biol Cell 17:4925-35
Li, Jie; Zhou, Lisa; Tran, Hoang T et al. (2006) Overexpression of laminin-8 in human dermal microvascular endothelial cells promotes angiogenesis-related functions. J Invest Dermatol 126:432-40
Gonzalez-Quevedo, Rosa; Shoffer, Marina; Horng, Lily et al. (2005) Receptor tyrosine phosphatase-dependent cytoskeletal remodeling by the hedgehog-responsive gene MIM/BEG4. J Cell Biol 168:453-63
Yant, Stephen R; Wu, Xiaolin; Huang, Yong et al. (2005) High-resolution genome-wide mapping of transposon integration in mammals. Mol Cell Biol 25:2085-94
Ortiz-Urda, Susana; Garcia, John; Green, Cheryl L et al. (2005) Type VII collagen is required for Ras-driven human epidermal tumorigenesis. Science 307:1773-6
Arbiser, Jack L; Fan, Chun-Yang; Su, Xiaobo et al. (2004) Involvement of p53 and p16 tumor suppressor genes in recessive dystrophic epidermolysis bullosa-associated squamous cell carcinoma. J Invest Dermatol 123:788-90

Showing the most recent 10 out of 58 publications