Biology; Chemistry; Endothelium, Vascular; Organic Chemicals; Pharmacology; Regeneration; Wound Healing; Keratinocytes; Endothelial Cells; Wnt Signaling Pathway
Postnatal Development & Regeneration
Stem Cell Center, Yale: Stem Cells and Tissue Repair
We use a combination of biochemistry, molecular biology, and bio-organicchemistry to explore different aspects of developmental and cell biology. Different projects include 1) the exploration of salamander limb regeneration with the goal of applying this knowledge to the possibility of mammalian regeneration, and 2) the exploration of how biologically active compounds from nature work in order to identify new probes for cell biology as well as identify novel drug targets.
Specialized Terms: Limb regeneration; wound epidermis; blastema; natural product mode of action studies; angiogenesis; Wnt signaling; protein turnover; protein kinase regulation
Extensive Research Description
Among tetrapods, urodele amphibians such as Mexican salamanders (aka axolotls) have the unique ability to regenerate missing lens, tails, jaws, and tails. This epimorphic phenomenon begins with the closure of the wound via migration of the existing surrounding epidermis, which over the course of several days becomes an unique structure known as the Regenerative Epidermis (RE). The RE is required for regeneration since its removal prevents limb regrowth; however, it is unclear if the RE plays a 'permissive' or an 'inductive' role in the generation of the undifferentiated cell mass (aka blastema) that accumulates under the RE. We have recently identified 124 genes that are expressed 1.5x more highly in the RE relative to non-regenerating, wound healing epidermis. We are currently exploring several of these secreted and membrane-bound proteins to determine if they play an inductive role in cellular dedifferentiation (i.e., blastema formation). If so, this induction of adult dedifferentiated cells could be considered a natural analogy to the generation of iPS cells in cell culture.In addition to our interest in regenerative medicine, we are also developing novel reagents, which will allow us to explore new areas in cell biology. This 'chemical genetic' approach uses biologically active natural products and de novo designed small molecules to identify critical components in intracellular processes. In the past few years, our efforts have focused on anti-angiogenic, antitumor and anti-inflammatory natural products. More recently, we have explored the use of small molecules to control intracellular protein levels, either by inhibiting their degradation or by inducing their proteolysis via the 26S proteasome. A goal of this research is to develop novel methodologies that would allow for small molecule control of the 'undruggable proteome'.Salamander Limb Regeneration: We studying the role of regeneration-specific genes in inducing cellular dedifferentiation.
Inducing Protein Turnover: We are developing new methodologies to use cell-permeable small molecules as inducers of targeted protein degradation.
- Hines J, Gough JD, Corson TW, Crews CM. Posttranslational protein knockdown coupled to receptor tyrosine kinase activation with phosphoPROTACs. Proc Natl Acad Sci U S A. (2013), May 28; 110(22):8942-7.
- Neklesa TK, Crews CM. Chemical biology: Greasy tags for protein removal. Nature. (2012) Jul 18;487(7407):308-9.
- Holman EC, Campbell LJ, and CM Crews. Microarray analysis of microRNA expression during axolotl limb regeneration. (2012) PLoS ONE doi/10.1371/journal.pone.0041804
- Sundberg TB, Darricarrere N, Griesbach H, Slusarski DC and CM Crews. (2011) Wnt Planar Cell Polarity Signaling Disruption by Aberrant Accumulation of the MetAP-2 Substrate Rab37. Chemistry & Biology, 18(10):1300-11
- Neklesa TK, Tae H-S, Schneekloth AR, Stulberg MJ, Corson TW, Sundberg TB, Holley SA, and CM Crews. (2011) Small-molecule hydrophobic tagging-induced degradation of HaloTag fusion proteins. Nature Chemical Biology, 7(8):538-43
- Campbell LJ, Suárez-Castillo EC, Ortiz-Zuazaga H, Knapp D, Tanaka EM, CM Crews. (2011) Gene expression profile of the regeneration epithelium during axolotl limb regeneration. Developmental Dynamics. 2011 240 (7):1826-40.
- A genetic interaction network of five genes for human polycystic kidney and liver diseases defines polycystin-1 as the central determinant of cyst formation. Fedeles SV, Tian X, Gallagher AR, Mitobe M, Nishio S, Lee SH, Cai Y, Geng L, CM Crews, Somlo S (2011). A genetic interaction network of five genes for human polycystic kidney and liver diseases defines polycystin-1 as the central determinant of cyst formation.
- Crews CM. (2010) Targeting the Undruggable Proteome: The Small Molecules of My Dreams. Chemistry & Biology, 17:551-555.
- Ju R, Cirone P, Lin S, Griesbach H, Slusarski DC and CM Crews. (2010) Activation of the Planar Cell Polarity Formin DAAM1 Leads to Inhibition of Endothelial Cell Proliferation, Migration and Angiogenesis, Proc. Nat. Acad. Sci, 107(15):6906-11.
- Raina K and CM Crews. (2010) Chemical Inducers of Targeted Protein Degradation. J. Biol. Chem, 285(15):11057-60.
- Campbell, LJ and CM Crews. (2008) Wound Epidermis Formation and Function in Urodele Amphibian Limb Regeneration. Cellular and Molecular Life Sciences Jan;65(1):73-9
- Leuenroth, SJ, N Bencivenga, P Igarishi, S Somlo, CM Crews. (2008) Triptolide Reduces Cystogenesis in a Model of Autosomal Dominant Polycystic Kidney Disease. J. Am. Soc. Nephrology 19:1659-1662.
- Molineaux, C, and CM Crews. (2008) Proteasome Inhibitors in Cancer Chemotherapy, Cancer: Principles and Practice of Oncology, 8th Ed. V.T DeVita, T.S. Lawrence, S.A. Rosenberg, editors. Chapter 25., pp.486-490.
- Leuenroth, S.J., et al. (2007). Triptolide is a traditional Chinese medicine-derived inhibitor of Polycystic Kidney Disease. Proc. Natl. Acad. Sci. (USA) 104:4380-4394.
- Yeh, J.R., et al. (2006). Targeted gene disruption of Methionine Aminopeptidase 2 Results in an embryonic gastrulation defect and endothelial cell growth arrest. Proc. Natl. Acad. Sci. (USA) 103(27):10379-84.