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Jordan Kreidberg, MD, PhD
Associate Professor of Pediatrics
Harvard Medical School/Children’s Hospital
Department of Nephrology


 

MRRC Project(s)

R01 DK50118
WT-1 in Organogenesis and Tumorigenesis

R01 DK 5704-01
Integrins in Kidney Epithelial Cell Morphology

The defined phenotype of the original WT-1 mutation, combined with a detailed knowledge of WT-1 action in vitro, provides an opportunity to genetically dissect at an organismic level how transcription factors regulate developmental processes in mammals. The first project proposes to undertake a more detailed mutagenic analysis to further our understanding of WT-1 function during development.

The interaction of epithelial cells with the basement membrane is mediated by integrin receptors for the extracellular matrix (ECM). Integrins are expressed on the basal membranes of epithelial cells and are believed to be the major class of receptors adhering cells to the basement membrane. The second project is an investigation of the molecular determinants by which the a3b1 integrin maintains epithelial morphology. Mice deficient in this integrin exhibit a disturbance in cerebral cortical neuronal development, perhaps affecting neuronal migration.

Research Description

Major Results

Organogenesis involves a programmed pattern of gene expression which controls cellular differentiation and morphogenesis. The developing kidney is one of the best characterized models for the study of the molecular control of organogenesis. The WT-1 gene, originally identified as a tumor suppressor gene associated with Wilms' tumors. a tumor of the kidney which occurs during childhood, is required during the inductive events at the beginning of kidney formation, and is also required for gonad formation. WT-1 also is expressed in the central nervous system and is likely to be involved in neural development. WT-1 is a zinc finger transcription factor with a proline-glutamine rich trans-regulatory region.

In the first project, novel gene targeting approaches will be used to express mutated forms of WT-1 from the WT-1 genomic locus. Abnormal developmental phenotypes obtained will be interpreted based on morphological changes, alterations in expression of other known markers of organ development, and on previous studies of WT-1 function in vitro, where similar WT-1 mutations have altered its action as a transcriptional regulator. Embryos carrying mutations in the WT-1 gene will be analyzed by in situ hybridization, immunohistology, and organ culture. RNA subtraction libraries will be constructed to find novel genes expressed in organ development.

In the second project, by deriving immortalized epithelial cell lines from wild type and a3b1 integrin-deficient renal collecting ducts we have established a system for studying how integrins maintain epithelial morphology. Our studies on mice deficient in a3b1 integrin have indicated that this receptor has important functions during the development of the kidneys, lungs, skin, and brain. In wild type cells a normal cortical cytoskeleton is present. In contrast a3b1 integrin-deficient cells assemble actin stress fibers. Our present results also indicate that a3b1 integrin is required for cadherin:catenin complexes to maintain their association with the cytoskeleton. Our proposed experiments will test the hypothesis that Rho family GTPases act as downstream effectors of a3b1 integrin to mediate cadherin:catenin mediated organization of the cortical cytoskeleton in epithelial cells. We will also test the hypothesis that the pattern of cytoskeletal assembly is determined by the integrin repertoire. These hypotheses will be tested by (1) analyzing downstream effectors of Rho family GTPases in epithelial cell lines from wild type and a3b1 integrin-deficient kidney collecting ducts; (2) analyzing cadherin:catenin:cytoskeletal interactions in cell lines transfected different with different integrin "-subunits; (3) assessing cadherin clustering in wild type and a3b1 integrin-deficient cells, and (4) determining the effects of b-catenin mutants on cytoskeletal assembly in wild type and a3b1 integrin-deficient cells.

Publications

Birk OS, Casiano DE, Wassif CA, Cogliati T, Zhao L, Zhao Y, Grinberg A, Huang S, Kreidberg JA, Parker KL, Porter FD, Westphal H. The LIM homebox gene Lhx9 is essential for mouse gonad formaton. Nature 2000;403:909.

DiPersio CM, Shao M, Di Costanzo L, Kreidberg JA, Hynes RO. Immortalized mouse keratinocytes acquire a3b1 integrin-dependent produ ction of MMP9/gelatinase B. J Cell Science 2000;113:2909-2921.

Dulabon L, Olson EC, Taglienti M, Eisenhuth S, McGrath B, Walsh CA, Kreidberg JA, Anton ES. Reelin binds a3b1 integrin and inhibits neuronal migration. Neuron 2000;27:33-44.

DiPersio CM, Neut R, Georges-Labousesse E, Kreidberg JA, Sonnenberg A, Hynes RO. a3b1 and a6b4 integrin receptors for laminin-5 are not essential for epidermal morphogenesis and homeostasis during skin development. J Cell Science 2000;113:3051.

Kreidberg JA. Functions of a3b1 integrin. Curr Opin Cell Biol 2000;12(5):548-53.

Kreidberg JA, Symons JM. Integrins in kidney development, function, and disease. Am J Physiol Renal Physiol 2000;279(2):F233-42.

Menko AS, Kreidberg JA, Ryan TT, van Bockstaele E, Kukuruzinska MA. Loss of a3b1 integrin results in an altered differentiation program in the mouse submandibular gland. Developmental Dynamics 2001;220:337-349.

Klinowska TCM, Alexander CM, Georges-Labouesse E, Van der Neut R, Kreidberg JA, Jones CJP, Sonnenberg A, Streuli CH. Epithelial development and differentiation in the mammary gland is not dependent on a3 or a6 integrin subunits. Developmental Biology 2001;233:449-467.

Lin Y, Liu A, Zhang S, Ruusunen T, Kreidberg JA, Peltoketo H, Drummon I, Vainio S. Induction of ureter branching as a response to Wnt-2b signaling during early kidney organogenesis. Developmental Dynamics 2001;222:26-39.

Wei Y, Eble JA, Wang Z, Kreidberg JA, Chapman HA. Urokinase receptors promote b1 integrin function through interaction with integrin a3b1. Molec Biol Cell 2001;12:2957-2986.

Zent R, Bush KT, Pohl ML, Quaranta V, Koshikawa N, Wang Z, Kreidberg JA, Sakurai H, Stuart RO, Nigam SK. Involvement of laminin binding integrins and Laminin-5 in branching morphogenesis of the ureteric bud during kidney development. Developmental Biology 2001;238:289-302.

Natoli TA, Liu J, Eremina V, Hodgens K, Li C, Hamano Y, Mundel P, Kalluri R, Miner JH, Quaggin SE, Kreidberg JA. A mutant form of the Wilms' tumor suppressor gene WT1 observed in Denys-Drash syndrome interferes with glomerular capillary development. J Am Soc Nephrol 2002;13(8):2058-67.

Menko AS, Zhang L, Schiano F, Kreidberg JA, Kukuruzinska MA. Regulation of cadherin junctions during mouse submandibular gland development. Dev Dyn 2002;224(3):321-33. 15. Kreidberg JA. Applications of adhesion molecule gene knockout cell lines. Methods Cell Biol 2002;69:309-24.

Natoli TA, McDonald A, Alberta JA, Taglienti ME, Housman DE, Kreidberg JA. A mammal-specific exon of WT1 is not required for development or fertility. Mol Cell Biol 2002;22(12):4433-8.

Yang X, Claas C, Kraeft SK, Chen LB, Wang Z, Kreidberg JA, Hemler ME. Palmitoylation of tetraspanin proteins: modulation of CD151 lateral interactions, subcellular distribution, and integrin-dependent cell morphology. Mol Biol Cell 2002;13(3):767-81.

Kreidberg J. Kidneys and sex, the Wilms' tumor connection. Pediatr Res 2002;51(2):128.

Chattopadhyay N, Wang Z, Ashman LK, Brady-Kalnay SM, Kreidberg JA. alpha3beta1 integrin-CD151, a component of the cadherin-catenin complex, regulates PTPmu expression and cell-cell adhesion. J Cell Biol 2003;163(6):1351-62.

Zhang F, Tom CC, Kugler MC, Ching TT, Kreidberg JA, Wei Y, Chapman HA. Distinct ligand binding sites in integrin alpha3beta1 regulate matrix adhesion and cell-cell contact. J Cell Biol 2003;163(1):177-88.

Zhang H, Palmer R, Gao X, Kreidberg J, Gerald W, Hsiao L, Jensen RV, Gullans SR, Haber DA. Transcriptional activation of placental growth factor by the forkhead/winged helix transcription factor FoxD1. Curr Biol 2003;13(18):1625-9. 22. Kreidberg JA. Podocyte differentiation and glomerulogenesis. J Am Soc Nephrol 2003;14(3):806-14.

He ZY, Brakebusch C, Fassler R, Kreidberg JA, Primakoff P, Myles DG. None of the integrins known to be present on the mouse egg or to be ADAM receptors are essential for sperm-egg binding and fusion. Dev Biol 2003;254(2):226-37.

Alberta JA, Springett GM, Rayburn H, Natoli TA, Loring J, Kreidberg JA, Housman D. Role of the WT1 tumor suppressor in murine hematopoiesis. Blood 2003;101(7):2570-4.

Natoli TA, Alberta JA, Bortvin A, Taglienti M, Menke D, Loring J, Jaenisch R, Page D, Housman DE, Kreidberg JA. WT1 is required cell-autonomously for the development of all gonadal cell lineages, including the germ cell to gonocyte transition. Dev Biol 2004, in press.

See Dr. Kreidberg's publications via PubMed

Contact Information

E-mail: Jordan Kreidberg, MD, PhD
Associate Professor of Pediatrics
Harvard Medical School/Children’s Hospital
Department of Nephrology