TY - JOUR
T1 - Unbiased Selection of Peptide-Peptoid Hybrids Specific for Lung Cancer Compared to Normal Lung Epithelial Cells
AU - Matharage, Jaya M.
AU - Minna, John D.
AU - Brekken, Rolf A.
AU - Udugamasooriya, D. Gomika
N1 - Funding Information:
We thank M. Peyton and J. Larsen at University of Texas Southwestern Medical Center (UTSW) for their initial help in establishing cancer and normal cell lines as well as important discussions. We also thank L. Girard at UTSW for confirming the genetic background of HCC4017 and HBEC30KT cell lines and Proteomics Core Facility at UTSW for conducting Edman Sequencing. We also thank K. Brown at UTSW for providing access to plater reader and MALDI mass spectrometer. This work was supported by National Cancer Institute (NCI) at National Institutes of Health (NIH) grant R01CA175779 and U01 CA176284, Cancer Prevention and Research Institute of Texas Grant RP130258 and RP110708, Lung Cancer SPORE P50CA70907 grant, and funding from University of Texas Southwestern Medical Center.
Publisher Copyright:
© 2015 American Chemical Society.
PY - 2015/12/18
Y1 - 2015/12/18
N2 - To develop widely applicable diagnostic and potentially therapeutic approaches overcoming protein heterogeneity in human cancer, we have developed a technology to unbiasedly select high specificity compound(s) that bind any biomolecule (e.g., proteins, lipids, carbohydrates) presented on the cancer cell surface but not on normal cells. We utilized a peptidomimetic based on-bead two-color (OBTC) combinatorial cell screen that can detect differences between two cell surfaces at high accuracy by looking for beads (where each bead in the library had one peptide-peptoid hybrid on the surface) that only bound cancer but not normal cells. We screened a library of 393216 compounds targeting HCC4017 lung adenocarcinoma cells (labeled in red) in the presence of HBEC30KT normal bronchial epithelial cells (labeled in green) derived from the same tissue of the same patient. This screen identified a peptide-peptoid hybrid called PPS1 which displayed high specific binding for HCC4017 cancer cells over HBEC30KT cells. Specificity was validated through on-bead, ELISA-like and magnetic bead pulldown studies, while a scrambled version of PPS1 did not show any binding. Of interest, the simple dimeric version (PPS1D1) displayed cytotoxic activity on HCC4017 cells, but not on normal HBEC30KT cells. PPS1D1 also strongly accumulated in HCC4017 lung cancer xenografts in mice over control constructs. We conclude that such combinatorial screens using tumor and normal cells from the same patient have significant potential to develop new reagents for cancer biology, diagnosis, and potentially therapy.
AB - To develop widely applicable diagnostic and potentially therapeutic approaches overcoming protein heterogeneity in human cancer, we have developed a technology to unbiasedly select high specificity compound(s) that bind any biomolecule (e.g., proteins, lipids, carbohydrates) presented on the cancer cell surface but not on normal cells. We utilized a peptidomimetic based on-bead two-color (OBTC) combinatorial cell screen that can detect differences between two cell surfaces at high accuracy by looking for beads (where each bead in the library had one peptide-peptoid hybrid on the surface) that only bound cancer but not normal cells. We screened a library of 393216 compounds targeting HCC4017 lung adenocarcinoma cells (labeled in red) in the presence of HBEC30KT normal bronchial epithelial cells (labeled in green) derived from the same tissue of the same patient. This screen identified a peptide-peptoid hybrid called PPS1 which displayed high specific binding for HCC4017 cancer cells over HBEC30KT cells. Specificity was validated through on-bead, ELISA-like and magnetic bead pulldown studies, while a scrambled version of PPS1 did not show any binding. Of interest, the simple dimeric version (PPS1D1) displayed cytotoxic activity on HCC4017 cells, but not on normal HBEC30KT cells. PPS1D1 also strongly accumulated in HCC4017 lung cancer xenografts in mice over control constructs. We conclude that such combinatorial screens using tumor and normal cells from the same patient have significant potential to develop new reagents for cancer biology, diagnosis, and potentially therapy.
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U2 - 10.1021/acschembio.5b00592
DO - 10.1021/acschembio.5b00592
M3 - Article
C2 - 26509598
AN - SCOPUS:84952837685
VL - 10
SP - 2891
EP - 2899
JO - ACS Chemical Biology
JF - ACS Chemical Biology
SN - 1554-8929
IS - 12
ER -