TY - JOUR
T1 - Delivery of Targeted Co(III)-DNA Inhibitors of Gli Proteins to Disrupt Hedgehog Signaling
AU - Dukes, Meghan W.
AU - Bajema, Elizabeth A.
AU - Whittemore, Tyler J.
AU - Holmgren, Robert A.
AU - Meade, Thomas J.
N1 - Funding Information:
This work was supported through the National Institute of Health Grant Numbers 1F31CA236175 (M.W.D.), 5F31CA186761 (E.A.B.), T32CA186897 (T.J.W.), and R01NS115571. The authors thank Professor Tony Oro for providing ASZ and BSZ cell lines, Christopher Brue for assistance in AFM imaging, and Timothy Goetjen for assistance with TGA. This work used the Keck-II facility of Northwestern University’s NUANCE Center, which has received support from the SHyNE Resource (NSF ECCS-2025633), the IIN, and Northwestern’s MRSEC program (NSF DMR-1720139). This work also used the IMSERC Physical Characterization facility at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-2025633) and Northwestern University. Microscopy was performed at the Biological Imaging Facility at Northwestern University (RRID: SCR_017767), graciously supported by the Chemistry for Life Processes Institute, the NU Office for Research, and the Department of Molecular Biosciences. Metal analysis was performed at the Northwestern University Quantitative Bio-element Imaging Center generously supported by NASA Ames Research Center NNA06CB93G.
Funding Information:
This work was supported through the National Institute of Health Grant Numbers 1F31CA236175 (M.W.D.), 5F31CA186761 (E.A.B.) T32CA186897 (T.J.W.), and R01NS115571. The authors thank Professor Tony Oro for providing ASZ and BSZ cell lines, Christopher Brue for assistance in AFM imaging, and Timothy Goetjen for assistance with TGA. This work used the Keck-II facility of Northwestern University?s NUANCE Center, which has received support from the SHyNE Resource (NSF ECCS-2025633), the IIN, and Northwestern?s MRSEC program (NSF DMR-1720139). This work also used the IMSERC Physical Characterization facility at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-2025633) and Northwestern University. Microscopy was performed at the Biological Imaging Facility at Northwestern University (RRID: SCR_017767), graciously supported by the Chemistry for Life Processes Institute, the NU Office for Research, and the Department of Molecular Biosciences. Metal analysis was performed at the Northwestern University Quantitative Bio-element Imaging Center generously supported by NASA Ames Research Center NNA06CB93G.
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/4/20
Y1 - 2022/4/20
N2 - The Hedgehog (Hh) signaling pathway is integral for embryonic development and normal cell maintenance. However, aberrant expression of the Hh pathway is recognized as the oncogenic driving force for basal cell carcinoma (BCC). Current chemotherapeutic treatments that inhibit Hh signaling allow treatment of only locally advanced and metastatic BCCs via inhibition of the transmembrane protein, smoothened. It is further recognized that downstream mutations often lead to chemoresistant tumor recurrence. The Gli proteins are the ultimate regulators of Hh signaling and belong to a family of Cys2His2zinc finger transcription factors (ZnFTFs) that we have shown can be irreversibly inhibited by a series of cobalt(III) Schiff base-DNA (CoSB-DNA) conjugates. However, a significant challenge is the delivery of CoSB-DNA complexes in mammalian tissues. Herein, we report a polyethyleneimine-functionalized graphene oxide nanoconjugate (GOPEI) that delivers CoGli, a CoSB-DNA complex that targets Gli specifically. We describe the characterization of the surface functionalization of GOPEI and accumulation in ASZ murine BCC cells via confocal microscopy and inductively coupled plasma-mass spectrometry (ICP-MS). Lysosomal escape of CoGli is further confirmed by confocal microscopy. We report the successful targeting of Gli by CoGli and a 17-fold improvement in potency over small-molecule Gli inhibitor GANT-61 in inhibiting Hh-driven migration of ASZ murine BCC cells. This study provides a promising starting point for further investigating CoGli inhibitors of Hh signaling in developed mammalian tissues.
AB - The Hedgehog (Hh) signaling pathway is integral for embryonic development and normal cell maintenance. However, aberrant expression of the Hh pathway is recognized as the oncogenic driving force for basal cell carcinoma (BCC). Current chemotherapeutic treatments that inhibit Hh signaling allow treatment of only locally advanced and metastatic BCCs via inhibition of the transmembrane protein, smoothened. It is further recognized that downstream mutations often lead to chemoresistant tumor recurrence. The Gli proteins are the ultimate regulators of Hh signaling and belong to a family of Cys2His2zinc finger transcription factors (ZnFTFs) that we have shown can be irreversibly inhibited by a series of cobalt(III) Schiff base-DNA (CoSB-DNA) conjugates. However, a significant challenge is the delivery of CoSB-DNA complexes in mammalian tissues. Herein, we report a polyethyleneimine-functionalized graphene oxide nanoconjugate (GOPEI) that delivers CoGli, a CoSB-DNA complex that targets Gli specifically. We describe the characterization of the surface functionalization of GOPEI and accumulation in ASZ murine BCC cells via confocal microscopy and inductively coupled plasma-mass spectrometry (ICP-MS). Lysosomal escape of CoGli is further confirmed by confocal microscopy. We report the successful targeting of Gli by CoGli and a 17-fold improvement in potency over small-molecule Gli inhibitor GANT-61 in inhibiting Hh-driven migration of ASZ murine BCC cells. This study provides a promising starting point for further investigating CoGli inhibitors of Hh signaling in developed mammalian tissues.
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U2 - 10.1021/acs.bioconjchem.2c00063
DO - 10.1021/acs.bioconjchem.2c00063
M3 - Article
C2 - 35271256
AN - SCOPUS:85126521377
SN - 1043-1802
VL - 33
SP - 643
EP - 653
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 4
ER -