Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells
BackgroundThe insulin-like growth factor 1 receptor (IGF-1R) and the thyrotropin receptor (TSH-R) are expressed on orbital cells and thyrocytes. These receptors are targeted in autoimmune-induced thyroid eye disease (TED). Effective therapeutic treatment of TED inhibits activation of the IGF-1R/TSH-...
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Frontiers Media S.A.
2024-12-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2024.1488220/full |
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| author | Maximilian Luffy Anna-Lena Ganz Stefanie Wagner Jan Wolf Julian Ropertz Ryan Zeidan Jeffrey D. Kent Raymond S. Douglas George J. Kahaly |
| author_facet | Maximilian Luffy Anna-Lena Ganz Stefanie Wagner Jan Wolf Julian Ropertz Ryan Zeidan Jeffrey D. Kent Raymond S. Douglas George J. Kahaly |
| author_sort | Maximilian Luffy |
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| description | BackgroundThe insulin-like growth factor 1 receptor (IGF-1R) and the thyrotropin receptor (TSH-R) are expressed on orbital cells and thyrocytes. These receptors are targeted in autoimmune-induced thyroid eye disease (TED). Effective therapeutic treatment of TED inhibits activation of the IGF-1R/TSH-R complex.MethodsThe inhibitory effect on cell proliferation of a small molecule targeting IGF-1R phosphorylation (Linsitinib) was investigated in an IGF-1R expressing cell line and a Chinese Hamster Ovary (CHO) cell line overexpressing TSH-R. An IGF-1R monoclonal antibody antagonist, Teprotumumab served as control. Both cell lines were plated in a 96-well format and treated with both compounds for 24 hours. After addition of tetrazolium, absorbance was measured. The apoptosis marker caspase-3/7 activity was measured. The half-maximal inhibitory concentration (IC50) of TSH-R-Ab induced stimulation (stimulatory monoclonal antibody, mAb, M22) of the TSH-R cell line was evaluated with a cell-based bioassay for blocking TSH-R-Ab. Cells were treated with ten rising concentrations of either Linsitinib, Linsitinib + Metformin, Teprotumumab, or a blocking TSH-R mAb (K1-70).ResultsLinsitinib strongly inhibited the proliferation of both cell lines at several concentrations: 31,612.5 ng/mL (IGF-1R cell line -78%, P=0.0031, TSH-R cell line -75%, P=0.0059), and at 63,225 ng/mL (IGF-1R cell line -73%, P=0.0073, TSH-R cell line -73%, P=0.0108). Linsitinib induced apoptosis of both cell lines, both morphologically confirmed and with an increased caspase-3/7 activity at concentrations of 31,612.5 ng/mL (IGF-1R cell line P=0.0158, TSH-R cell line P=0.0048) and 63,225 ng/mL (IGF-1R cell line P=0.0005, TSH-R cell line P=0.0020). Linsitinib markedly inhibited proliferation of the IGF-1R cell line at all concentrations compared to Teprotumumab (P=0.0286). Teprotumumab inhibition was significant only at 15,806.25 ng/mL with the TSH-R cell line (-15%, P=0.0396). In addition, in the TSH-R-Ab blocking bioassay, Linsitinib and the tested compounds demonstrated strong inhibition across all ten dilutions (100%).ConclusionsLinsitinib effectively induces apoptosis and inhibits proliferation of both IGF-1R and TSH-R expressing target cells, therefore demonstrating its therapeutic potential to block the reported crosstalk of the two mediators in autoimmune TED. |
| format | Article |
| id | doaj-art-e92264e527924748b7fcf8e8d63e3420 |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2024-12-01 |
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| spelling | doaj-art-e92264e527924748b7fcf8e8d63e34202024-12-11T06:44:52ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-12-011510.3389/fimmu.2024.14882201488220Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cellsMaximilian Luffy0Anna-Lena Ganz1Stefanie Wagner2Jan Wolf3Julian Ropertz4Ryan Zeidan5Jeffrey D. Kent6Raymond S. Douglas7George J. Kahaly8Molecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg-University (JGU) Medical Center, Mainz, GermanyMolecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg-University (JGU) Medical Center, Mainz, GermanyMolecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg-University (JGU) Medical Center, Mainz, GermanyMolecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg-University (JGU) Medical Center, Mainz, GermanyMolecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg-University (JGU) Medical Center, Mainz, GermanySling Therapeutics, Ann Arbor, MI, United StatesSling Therapeutics, Ann Arbor, MI, United StatesSling Therapeutics, Ann Arbor, MI, United StatesMolecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg-University (JGU) Medical Center, Mainz, GermanyBackgroundThe insulin-like growth factor 1 receptor (IGF-1R) and the thyrotropin receptor (TSH-R) are expressed on orbital cells and thyrocytes. These receptors are targeted in autoimmune-induced thyroid eye disease (TED). Effective therapeutic treatment of TED inhibits activation of the IGF-1R/TSH-R complex.MethodsThe inhibitory effect on cell proliferation of a small molecule targeting IGF-1R phosphorylation (Linsitinib) was investigated in an IGF-1R expressing cell line and a Chinese Hamster Ovary (CHO) cell line overexpressing TSH-R. An IGF-1R monoclonal antibody antagonist, Teprotumumab served as control. Both cell lines were plated in a 96-well format and treated with both compounds for 24 hours. After addition of tetrazolium, absorbance was measured. The apoptosis marker caspase-3/7 activity was measured. The half-maximal inhibitory concentration (IC50) of TSH-R-Ab induced stimulation (stimulatory monoclonal antibody, mAb, M22) of the TSH-R cell line was evaluated with a cell-based bioassay for blocking TSH-R-Ab. Cells were treated with ten rising concentrations of either Linsitinib, Linsitinib + Metformin, Teprotumumab, or a blocking TSH-R mAb (K1-70).ResultsLinsitinib strongly inhibited the proliferation of both cell lines at several concentrations: 31,612.5 ng/mL (IGF-1R cell line -78%, P=0.0031, TSH-R cell line -75%, P=0.0059), and at 63,225 ng/mL (IGF-1R cell line -73%, P=0.0073, TSH-R cell line -73%, P=0.0108). Linsitinib induced apoptosis of both cell lines, both morphologically confirmed and with an increased caspase-3/7 activity at concentrations of 31,612.5 ng/mL (IGF-1R cell line P=0.0158, TSH-R cell line P=0.0048) and 63,225 ng/mL (IGF-1R cell line P=0.0005, TSH-R cell line P=0.0020). Linsitinib markedly inhibited proliferation of the IGF-1R cell line at all concentrations compared to Teprotumumab (P=0.0286). Teprotumumab inhibition was significant only at 15,806.25 ng/mL with the TSH-R cell line (-15%, P=0.0396). In addition, in the TSH-R-Ab blocking bioassay, Linsitinib and the tested compounds demonstrated strong inhibition across all ten dilutions (100%).ConclusionsLinsitinib effectively induces apoptosis and inhibits proliferation of both IGF-1R and TSH-R expressing target cells, therefore demonstrating its therapeutic potential to block the reported crosstalk of the two mediators in autoimmune TED.https://www.frontiersin.org/articles/10.3389/fimmu.2024.1488220/fullLinsitinibsmall molecule kinase inhibitorinsulin-like growth factor 1 receptorthyrotropin receptorthyroid eye diseaseapoptosis |
| spellingShingle | Maximilian Luffy Anna-Lena Ganz Stefanie Wagner Jan Wolf Julian Ropertz Ryan Zeidan Jeffrey D. Kent Raymond S. Douglas George J. Kahaly Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells Frontiers in Immunology Linsitinib small molecule kinase inhibitor insulin-like growth factor 1 receptor thyrotropin receptor thyroid eye disease apoptosis |
| title | Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells |
| title_full | Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells |
| title_fullStr | Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells |
| title_full_unstemmed | Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells |
| title_short | Linsitinib inhibits proliferation and induces apoptosis of both IGF-1R and TSH-R expressing cells |
| title_sort | linsitinib inhibits proliferation and induces apoptosis of both igf 1r and tsh r expressing cells |
| topic | Linsitinib small molecule kinase inhibitor insulin-like growth factor 1 receptor thyrotropin receptor thyroid eye disease apoptosis |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2024.1488220/full |
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