May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Identification of Small Mole
Author Affiliations & Notes
  • A. J. Johnson
    Scripps Research Institute, La Jolla, California
    Cell Biology,
  • M. I. Dorell
    Scripps Research Institute, La Jolla, California
    Cell Biology,
  • A. Shagini
    Scripps Research Institute, La Jolla, California
    Chemistry,
  • M. R. Ritter
    Scripps Research Institute, La Jolla, California
    Cell Biology,
  • W. Jin
    Scripps Research Institute, La Jolla, California
    Chemistry,
  • S. Moreno
    Scripps Research Institute, La Jolla, California
    Cell Biology,
  • D. L. Boger
    Scripps Research Institute, La Jolla, California
    Chemistry,
  • M. Friedlander
    Scripps Research Institute, La Jolla, California
    Cell Biology,
  • Footnotes
    Commercial Relationships A.J. Johnson, None; M.I. Dorell, None; A. Shagini, None; M.R. Ritter, None; W. Jin, None; S. Moreno, None; D.L. Boger, None; M. Friedlander, None.
  • Footnotes
    Support National Eye Institute Grant EY011254 (MF) and EY014174 (MF) , CIRM Post-doctoral Fellowship (MD), MacTel Foundation (MF), Scripps Fonseca/Mericos Fund (MF)
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 4103. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      A. J. Johnson, M. I. Dorell, A. Shagini, M. R. Ritter, W. Jin, S. Moreno, D. L. Boger, M. Friedlander; Identification of Small Mole. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4103.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose:: Bone marrow-derived progenitor cells (BMCs) have been shown to target sites of neovascularization in the retina and exhibit vasculo- and neurotrophic rescue in retinal degenerative mouse models (Otani et al., 2002; Otani et al., 2004). They have also demonstrated potent rescue functions in models of ischemic retinopathy (Ritter et al., 2006). BMCs have been shown to differentially express the hyaluronic acid receptor CD44. Current methods of enriching the functional subpopulation of BMCs utilize differential expression levels of CD44; the functional subpopulation expresses high levels of CD44 (CD44Hi) while low level expressing cells (CD44Lo) are non-functional. The CD44Hi population of cells consists mainly of myeloid progenitors, while the majority of non-functional CD44Lo cells are erythroid or lymphoid in origin (Ritter et al., 2006). Together, Ter119 and CD45RB220 identify the majority of non-functional cells, binding the erythroid and lymphoid cells respectively. Thus Ter119 and CD45RB220 depleted populations of cells would represent a functional population of cells free of bound purification agents such as antibodies or magnetic beads.

Methods:: We have used an in vitro, cell based selection assay to screen a peptidomimetic library of 60,000 molecules with modifiable side chains (Boger et al., 2003) to identify small molecules that efficiently bind to CD44Lo -expressing cells, for use as a negative selection tool. We have screened this library and identified compounds that competitively inhibit the binding of anti-Ter119 or anti-CD45R-B220 to CD44 Lo BMCs.

Results:: Candidate molecules that inhibit binding of either Ter-119 or CD45R-B220 antibody to CD44Lo-expressing cells have been identified from a library of non-peptidic chemicals. Of 1000 candidate molecules screened, 35 blocked antibody binding to the BMCs. 6 of these displayed efficient dose-dependant inhibition of antibody binding. Interestingly, the most promising candidates exhibited R groups that were similar in both orientation and structure.

Conclusions:: We have developed a small-molecule based selection procedure. From the initial screen, we have identified 6 candidates that exhibit reproducible, significant antibody inhibition in a dose-dependant fashion. With appropriate chemical modifications, this could (1) eliminate the need to use immunoglobulins and (2) provide a readily modifiable chemical structure that could negatively select subpopulations of BMCs with retinal vascular rescue potential.

Keywords: retinal neovascularization • retinal development • neuroprotection 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×