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Children’s Hospital Los Angeles

3 ARTICLES PUBLISHED IN JoVE

Cancer Research

Aqueous Humor as a Liquid Biopsy for Retinoblastoma: Clear Corneal Paracentesis and Genomic Analysis

Mary E. Kim^1,2, Liya Xu^1,2,3,11, Rishvanth K. Prabakar⁴, Lishuang Shen⁵, Chen-Ching Peng^1,3, Peter Kuhn^3,6,7,8, Xiaowu Gai^5,9, James Hicks^3,6,10, Jesse L. Berry^1,2,6,11

¹The Vision Center, Children’s Hospital Los Angeles, ²USC Roski Eye Institute, Keck School of Medicine of the University of Southern California, ³Department of Biological Sciences, Dornsife College of Letters, Arts, and Sciences, University of Southern California, ⁴Department of Molecular and Computational Biology, University of Southern California, ⁵Center for Personalized Medicine, Department of Pathology and Laboratory Medicine, Children’s Hospital Los Angeles, ⁶Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, ⁷Department of Aerospace and Mechanical Engineering, Viterbi School of Engineering, University of Southern California, ⁸Department of Biomedical Engineering, Viterbi School of Engineering, University of Southern California, ⁹Department of Pathology and Laboratory Medicine, Keck School of Medicine of USC, ¹⁰Department of Biochemistry and Molecular Medicine, Keck School of Medicine, University of Southern California, ¹¹The Saban Research Institute, Children’s Hospital Los Angeles

The aqueous humor is a high-yield liquid biopsy for retinoblastoma, intraocular cancer that cannot be biopsied in vivo due to the risk of extraocular spread. Herein, a method for safely extracting aqueous humor via clear corneal paracentesis and steps for genomic analysis to identify prognostic biomarkers are presented.

Developmental Biology

Modeling Paracrine Noncanonical Wnt Signaling In Vitro

Omar Toubat¹, Jongkyu Choi¹, S. Ram Kumar^1,2,3

¹Department of Surgery, Keck School of Medicine of USC, ²Department of Pediatrics, Keck School of Medicine of USC, ³Heart Institute, Children’s Hospital Los Angeles

The present study outlines a highly reproducible and tractable method to study paracrine noncanonical Wnt signaling events in vitro. This protocol was applied to evaluate the impact of paracrine Wnt5a signaling in murine neural crest cells and myoblasts.

Biology

Obtaining High-Quality Cryosections of Whole Rabbit Eye

Erik A. Souverein¹, Aaron Nagiel^2,3,4, Ksenia Gnedeva^5,6

¹Keck School of Medicine, University of Southern California, ²The Vision Center, Department of Surgery, Children’s Hospital Los Angeles, ³The Saban Research Institute, Children’s Hospital Los Angeles, ⁴USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine, University of Southern California, ⁵Eli and Edythe Broad CIRM Center for Regenerative Medicine and Stem Cell Research, University of Southern California, ⁶Tina and Rick Caruso Department of Otolaryngology-Head and Neck Surgery, University of Southern California

This protocol describes a reliable method for obtaining high-quality cryosections of whole rabbit eyes. It details rabbit eye dissection, fixation, embedding, and sectioning procedures, which may be easily adapted for use in any study utilizing immunohistochemistry in larger eyes.