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NT5E binds OTUD4. 56 / 56
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"The heatmap presented in Supplemental Figure 10D displays the cross-correlation between the residues P264, I266, R273, and K274 that coordinate Z22, suggesting that Z22 binding to that specific pocket allosterically affects the conformation of the binding epitope, thus interfering with complex formation.To further understand the inhibitory effects of ST80 and Z22 on the regulation of CD73 stability, we performed a series of experiments to determine the optimal dose and treatment duration for ST80 and Z22 to disrupt the OTUD4/CD73 interaction."
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"The data together support a great potential of pharmacological targeting of the proteolytic interaction of OTUD4 and CD73 to mitigate tumor immune evasion and revive antitumor T cell immunity."
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"Pharmacological blockade of interaction of OTUD4 and CD73 promotes tumor immunogenicity and inhibits tumor progression in immune-cold breast cancer.."
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"To evaluate the importance of the physical interaction between OTUD4 and CD73 for tumor growth, we overexpressed both OTUD4 and CD73 (OTUD4 OE + CD73 ) or OTUD4 and CD73 with mutated interaction sites (OTUD4 OE + CD73 ) in EO771 cells."
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"ST80, a novel inhibitor that destroys the proteolytic interaction between OTUD4 and CD73, has the potential to enhance the efficacy of PD-L1 treatment in TNBC ( xref )."
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"However, we still observed the interaction between OTUD4 and CD73, although its proportion correlated with the decreased level of OTUD4 and CD73 proteins ( xref )."
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"Notably, OTUD4 overexpression heightened levels of membrane-bound CD73 in cancer cells."
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"In addition, ST80, a newly developed inhibitor, specifically disrupted proteolytic interaction between CD73 and OTUD4, leading to reinvigoration of cytotoxic CD8 + T cell activities."
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"More importantly, we have developed a pharmacologic inhibitor, ST80 that is potent to specifically inhibit the physical interaction of OTUD4 and CD73 to promote ubiquitylation of CD73, thereby causing CD73 degradation and revival of CD8 T cell function."
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"These findings indicate a therapeutic value of pharmacologically targeting the proteolytic interaction of OTUD4 and CD73 to overcome the resistance of immune checkpoint inhibitors (e.g., anti-PD-L1 antibodies), especially in high CD73-expressing, immunologically suppressive TNBC."
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"The data together support a great potential of pharmacological targeting of the proteolytic interaction of OTUD4 and CD73 to mitigate tumor immune evasion and revive antitumor T cell immunity.To validate the spatially correlated expression of OTUD4 and CD73 in tumor cells with immune signaling programs in the tumor microenvironment, we performed spatially indexed transcript profiling (GeoMx digital spatial profiling) of tumor tissue sections from 6 patients with TNBC."
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"Targeting a specific interaction between tumor OTUD4 and CD73 to efficiently destabilize CD73 is an effective strategy for TNBC treatment, especially when combined with PD-L1 blockade, even in the case with abundant expression of tumor CD73 and OTUD4."
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"Using gene pathway analysis (Kyoto Encyclopedia of Genes and Genomes [KEGG]/Reactome) ( xref ) based on spatial transcriptomes, we found Ubl conjugation, TGF-β receptor activity, and SMAD binding pathways pertaining to CD73 PTM modulation were particularly enriched in OTUD4 lo compared with OTUD4 hi tumor areas, further supporting potential protein interactions between OTUD4 and CD73 in tumor cells."
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"Subsequent immunoprecipitation assays consistently showed CD73 interacting with OTUD4 in TNBC cells, and in situ detection techniques showed the interaction occurred within the cytosol."
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"We further developed a pharmacological inhibitor, ST80, to resume CD73 proteolysis and restore capacity to elicit CD8 + interferon-γ-producing T cell responses through the blockade of the interaction between OTUD4 and CD73."
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"To validate the biochemical consequences as well as physiological relevance of interaction between OTUD4 and CD73, we performed a series of immunoprecipitation assays in the MDA-MB468 cells ( xref )."
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"To further validate the generality of the OTUD4/CD73 interaction, a series of immunoprecipitation assays across diverse cancer cell lines, including the nonsmall cell lung cancer cell HCC827, human ovarian cancer cell SKOV3, and human colorectal cancer cell HCT116, consistently showed endogenous interaction between CD73 and OTUD4 ( xref , A–D)."
38530357
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"More importantly, we have developed a pharmacologic inhibitor, ST80 that is potent to specifically inhibit the physical interaction of OTUD4 and CD73 to promote ubiquitylation of CD73, thereby causing CD73 degradation and revival of CD8 + T cell function."
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"To examine the cellular compartmentalization of the interaction between CD73 and OTUD4, we conducted cell lysate fractionation followed by Western blotting ( xref )."
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"An important therapeutic advance proposed by Zhu, Banerjee, and colleagues is the development of ST80, a pharmacologic inhibitor designed to target the physical interaction between OTUD4 and CD73 ( xref ) ( xref )."
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"In this context, targeting the protein-protein interactions between CD73 and its upstream deubiquitinase OTUD4 in destabilizing CD73 and reducing its cell surface abundance provides a therapeutic rationale."
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"As presented in xref , although CD73 (green) was observed both in membrane and cytosol and OTUD4 (red) was observed both in nucleus and cytosol, the interaction between CD73 and OTUD4, in the cytosol as determined by proximity ligation assay ( xref ), suggested a potential link between OTUD4-guided cytosolic deubiquitylation of CD73 and its subsequent displacement onto the cell membrane."
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"We implemented a triphase methodology to identify small molecules that could block the interaction between CD73 and OTUD4 ( xref )."
38530357
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"This inhibitor disrupted the interaction between OTUD4 and CD73, leading to the ubiquitylation and subsequent degradation of CD73."
38747288
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"Targeting a specific interaction between tumor OTUD4 and CD73 to efficiently destabilize CD73 is an effective strategy for TNBC treatment, especially when combined with PD-L1 blockade, even in the case with abundant expression of tumor CD73 and OTUD4."
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"Targeting the specific interaction between OTUD4 and CD73 offers a promising approach for TNBC treatment, especially in combination with immune checkpoint inhibitor therapy."
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"Mapping and structural modeling of the interaction between CD73 and OTUD4 by integrated use of experimental data and docking simulations followed by experimental validation."
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"We further narrowed down to amino acids stretching from 380 to 500 on OTUD4 that mediated the interaction between OTUD4 and CD73 ( xref , A and B)."
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"Given the significance of hydrophobic contacts at the CD73-OTUD4 interface, we focused on CD73 V300 and I301."
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"ST80, a novel inhibitor that destroys the proteolytic interaction between OTUD4 and CD73, has the potential to enhance the efficacy of PD-L1 treatment in TNBC (121)."
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"To further confirm if amino acids V300 and I301 on CD73 directly mediate its binding to OTUD4, the results depicted in xref and xref reveal a marked decrease in the interaction between mutant CD73 and OTUD4 when the amino acids V300 and I301 are mutated in both MDA-MB231 and MDA-MB468 cells."
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"As indicated in xref and xref , MDA-MB468-CD73 WT cells had a decrease in IFN-γ production and proliferation capacity of CD8 + T cell population than MDA-MB468 vehicle control cells, and the disruption of the CD73-OTUD4 interaction by mutating the V300/I301 sites restored the IFN-γ production and proliferation capacity of CD8 + T cell population comparable to those of the control vector."
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"The data demonstrate the specific interaction between CD73 and OTUD4, dictated through the V300 and I301 residues on CD73, plays a pivotal role in governing CD73 protein stability and subsequent adenosinergic effect, particularly on modulation of T cell activity."
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"As to the mode of action of Z22, the binding position shown in xref suggests that it acts as an allosteric inhibitor of the CD73-OTUD4 interaction."
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"We further developed a pharmacological inhibitor, ST80, to resume CD73 proteolysis and restore capacity to elicit CD8 interferon-γ-producing T cell responses through the blockade of the interaction between OTUD4 and CD73."
38530357
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"To validate the biochemical consequences as well as physiological relevance of interaction between OTUD4 and CD73, we performed a series of immunoprecipitation assays in the MDA-MB468 cells (26)."
38530357
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"To further validate the generality of the OTUD4/CD73 interaction, a series of immunoprecipitation assays across diverse cancer cell lines, including the nonsmall cell lung cancer cell HCC827, human ovarian cancer cell SKOV3, and human colorectal cancer cell HCT116, consistently showed endogenous interaction between CD73 and OTUD4 (Supplemental Figure 4, A–D)."
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"To evaluate the importance of the physical interaction between OTUD4 and CD73 for tumor growth, we overexpressed both OTUD4 and CD73 WT (OTUD4 OE + CD73 WT ) or OTUD4 and CD73 V300P/I301Q with mutated interaction sites (OTUD4 OE + CD73 V300P/I301Q ) in EO771 cells."
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"To examine the cellular compartmentalization of the interaction between CD73 and OTUD4, we conducted cell lysate fractionation followed by Western blotting (27)."
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"As presented in Supplemental Figure 4E, although CD73 (green) was observed both in membrane and cytosol and OTUD4 (red) was observed both in nucleus and cytosol, the interaction between CD73 and OTUD4, in the cytosol as determined by proximity ligation assay (Supplemental Figure 4F), suggested a potential link between OTUD4-guided cytosolic deubiquitylation of CD73 and its subsequent displacement onto the cell membrane."
38530357
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"Mapping and structural modeling of the interaction between CD73 and OTUD4 by integrated use of experimental data and docking simulations followed by experimental validation.."
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"We further narrowed down to amino acids stretching from 380 to 500 on OTUD4 that mediated the interaction between OTUD4 and CD73 (Supplemental Figure 6, A and B)."
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"To comprehensively address the mechanism by which the interplay of TRIM21-mediated ubiquitylation and OTUD4-guided deubiquitylation regulates CD73 proteolysis, we conducted structural modeling of the interaction between CD73, TRIM21, and OTUD4 by integrated use of experimental data and docking simulations followed by experimental validation."
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"The interface is further characterized by 14 hydrogen bonds and 2 salt bridges.To identify the key amino acids that drive the OTUD4/CD73 interaction, Supplemental Figure 6E details the types and strengths of inter-residue interactions."
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"Pharmacological blockade of interaction of OTUD4 and CD73 promotes tumor immunogenicity and inhibits tumor progression in immune-cold breast cancer."
38530357
reach
"An important therapeutic advance proposed by Zhu, Banerjee, and colleagues is the development of ST80, a pharmacologic inhibitor designed to target the physical interaction between OTUD4 and CD73 (Figure 1) (3)."
38747288
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"This inhibitor disrupted the interaction between OTUD4 and CD73, leading to the ubiquitylation and subsequent degradation of CD73."
38747288
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"To understand the mechanisms by which ST80 and Z22 disrupt the interaction between OTUD4 and CD73, we illustrated in xref that ST80 binds to a hydrophobic area defined by residues I71, P254, Y289, P307, I308, and L309."
38530357
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"In addition, ST80, a newly developed inhibitor, specifically disrupted proteolytic interaction between CD73 and OTUD4, leading to reinvigoration of cytotoxic CD8 + T cell activities."
38530357
sparser
"These findings indicate a therapeutic value of pharmacologically targeting the proteolytic interaction of OTUD4 and CD73 to overcome the resistance of immune checkpoint inhibitors (e.g., anti-PD-L1 antibodies), especially in high CD73-expressing, immunologically suppressive TNBC."
38530357
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"However, we still observed the interaction between OTUD4 and CD73, although its proportion correlated with the decreased level of OTUD4 and CD73 proteins (Supplemental Figure 8F)."
38530357
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"Development of a pharmacological inhibitor that blocks OTUD4/CD73 interaction in restoring tumor immune responses in immune-suppressive TNBCs.."
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"We next aimed to determine the therapeutic consequence of blocking the OTUD4/CD73 complex formation toward restoring tumor immune response in immune-suppressive TNBCs."
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"We implemented a triphase methodology to identify small molecules that could block the interaction between CD73 and OTUD4 (Supplemental Figure 9A)."
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"As shown in Figure 5, B and C, our systematic validation led to the identification of 2 compounds, ZINC000009345994 (ST80) and ZINC001336656227 (Z22), as the most potent inhibitors in suppressing CD73 protein abundance by blocking the OTUD4/CD73 interaction."
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"These results together suggest that both ST80 and Z22 are potential pharmacological inhibitors that specifically block OTUD4/CD73 interaction to destabilize CD73 protein.To understand the mechanisms by which ST80 and Z22 disrupt the interaction between OTUD4 and CD73, we illustrated in Figure 5F that ST80 binds to a hydrophobic area defined by residues I71, P254, Y289, P307, I308, and L309."
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