Cannabinoid receptor type 1 (CB1R) inhibits hypothalamic leptin signaling via β-arrestin1 in complex with TC-PTP and STAT3

Gergő Szanda; Tony Jourdan; Éva Wisniewski; Resat Cinar; Grzegorz Godlewski; Anikó Rajki; Jie Liu; Lee Chedester; Bence Szalai; András Dávid Tóth; Eszter Soltész-Katona; László Hunyady; Asuka Inoue; Viktória Bea Horváth; András Spät; Joseph Tam; George Kunos

doi:10.1016/j.isci.2023.107207

Cannabinoid receptor type 1 (CB₁R) inhibits hypothalamic leptin signaling via β-arrestin1 in complex with TC-PTP and STAT3

Gergő Szanda^*, Tony Jourdan, Éva Wisniewski, Resat Cinar, Grzegorz Godlewski, Anikó Rajki, Jie Liu, Lee Chedester, Bence Szalai, András Dávid Tóth, Eszter Soltész-Katona, László Hunyady, Asuka Inoue, Viktória Bea Horváth, András Spät, Joseph Tam, George Kunos^*

^*Corresponding author for this work

School of Pharmacy

Research output: Contribution to journal › Article › peer-review

Abstract

Molecular interactions between anorexigenic leptin and orexigenic endocannabinoids, although of great metabolic significance, are not well understood. We report here that hypothalamic STAT3 signaling in mice, initiated by physiological elevations of leptin, is diminished by agonists of the cannabinoid receptor 1 (CB₁R). Measurement of STAT3 activation by semi-automated confocal microscopy in cultured neurons revealed that this CB₁R-mediated inhibition requires both T cell protein tyrosine phosphatase (TC-PTP) and β-arrestin1 but is independent of changes in cAMP. Moreover, β-arrestin1 translocates to the nucleus upon CB₁R activation and binds both STAT3 and TC-PTP. Consistently, CB₁R activation failed to suppress leptin signaling in β-arrestin1 knockout mice in vivo, and in neural cells deficient in CB₁R, β-arrestin1 or TC-PTP. Altogether, CB₁R activation engages β-arrestin1 to coordinate the TC-PTP-mediated inhibition of the leptin-evoked neuronal STAT3 response. This mechanism may restrict the anorexigenic effects of leptin when hypothalamic endocannabinoid levels rise, as during fasting or in diet-induced obesity.

Original language	American English
Article number	107207
Journal	iScience
Volume	26
Issue number	7
DOIs	https://doi.org/10.1016/j.isci.2023.107207
State	Published - 21 Jul 2023

Bibliographical note

Funding Information:
This work was supported by the following grants: National Research, Development and Innovation Office grants NKFI-6/FK_124038 to G. Szanda, intramural NIH funds to G. Kunos and FK_ 18/128376 to É.W. (PI: Roland Csépányi-Kömi). The work was also funded by the Scientific and Innovation Fund of the Semmelweis University , Budapest, Hungary ( 26303/AOELT/2019 ; to G.S.) and the Eötvös Loránd Research Network (to ELKH-SE Laboratory of Molecular Physiology Research Group ). G. Szanda was supported by the János Bólyai Research Scholarship of the Hungarian Academy of Sciences . The high-content imaging system was procured using Hungarian National Competitiveness and Excellence Programme funds ( NVKP_16-1-2016-0039 ).

Funding Information:
We are indebted to Dr. Katalin Erdélyi (National Institute of Oncology, Budapest, Hungary), Dr. Pál Pacher (NIAAA, NIH, Rockville, MD, USA), Dr. Balázs Enyedi (Semmelweis University, Budapest, Hungary) and Dr. Zoltán Hegyi (Bio-Science, Budapest, Hungary) for technical help and valuable discussion. GT1-7 cells were kindly provided by Dr. Pamela L. Mellon (University of California, San Diego, CA, USA). The authors thank Dr. Gerhard Müller-Newen (University Hospital Aachen, Aachen, Germany) for generously providing the STAT3-CFP-YFP construct and Dr. Tamás Balla (NICHD, NIH, Bethesda, MD, USA) for his help in construct design and production. The CFP-tagged mouse leptin receptor clone was a generous gift from Dr. Arieh Gertler (The Hebrew University of Jerusalem, Rehovot, Israel). We are indebted to Dr. Erzsébet Ligeti and Dr. Roland Csépányi-Kömi (Semmelweis University, Budapest, Hungary) for providing resources for molecular biological work. This work was supported by the following grants: National Research, Development and Innovation Office grants NKFI-6/FK_124038 to G. Szanda, intramural NIH funds to G. Kunos and FK_18/128376 to É.W. (PI: Roland Csépányi-Kömi). The work was also funded by the Scientific and Innovation Fund of the Semmelweis University, Budapest, Hungary (26303/AOELT/2019; to G.S.) and the Eötvös Loránd Research Network (to ELKH-SE Laboratory of Molecular Physiology Research Group). G. Szanda was supported by the János Bólyai Research Scholarship of the Hungarian Academy of Sciences. The high-content imaging system was procured using Hungarian National Competitiveness and Excellence Programme funds (NVKP_16-1-2016-0039). The Servier Medical Art Collection (https://smart.servier.com/) was used for the preparation of the graphical abstract. Conceptualization G.S. and G.K.; Methodology G.S. T.J. J.T. R.C. and G.K.; Formal Analysis G.S. A.S. E.S-K. A.D.T. and G.K. Investigation G.S. T.J. R.C. G.G. A.R. J.L. L.C. B.S. E.S-K. A.D.T. V.B.H. and J.T.; Resources G.S. E.W. L.H. A.I. and G.K.; Writing – Original Draft G.S. and G.K.; Writing – Review & Editing G.S. T.J. A.S. A.D.T. J.T. and G.K. Visualization G.S.; Supervision G.S. and G.K.; Funding Acquisition G.S. L.H. and G.K. All authors had access to the manuscript and agreed with the final version. The authors declare no competing interest. We support inclusive, diverse and equitable conduct of research.

Publisher Copyright:
© 2023 The Author(s)

Keywords

Cell biology
Molecular biology

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10.1016/j.isci.2023.107207

Cite this

Szanda, G., Jourdan, T., Wisniewski, É., Cinar, R., Godlewski, G., Rajki, A., Liu, J., Chedester, L., Szalai, B., Tóth, A. D., Soltész-Katona, E., Hunyady, L., Inoue, A., Horváth, V. B., Spät, A., Tam, J., & Kunos, G. (2023). Cannabinoid receptor type 1 (CB₁R) inhibits hypothalamic leptin signaling via β-arrestin1 in complex with TC-PTP and STAT3. iScience, 26(7), Article 107207. https://doi.org/10.1016/j.isci.2023.107207

@article{e71fad3a943e410e93a5edff52092fa5,

title = "Cannabinoid receptor type 1 (CB1R) inhibits hypothalamic leptin signaling via β-arrestin1 in complex with TC-PTP and STAT3",

abstract = "Molecular interactions between anorexigenic leptin and orexigenic endocannabinoids, although of great metabolic significance, are not well understood. We report here that hypothalamic STAT3 signaling in mice, initiated by physiological elevations of leptin, is diminished by agonists of the cannabinoid receptor 1 (CB1R). Measurement of STAT3 activation by semi-automated confocal microscopy in cultured neurons revealed that this CB1R-mediated inhibition requires both T cell protein tyrosine phosphatase (TC-PTP) and β-arrestin1 but is independent of changes in cAMP. Moreover, β-arrestin1 translocates to the nucleus upon CB1R activation and binds both STAT3 and TC-PTP. Consistently, CB1R activation failed to suppress leptin signaling in β-arrestin1 knockout mice in vivo, and in neural cells deficient in CB1R, β-arrestin1 or TC-PTP. Altogether, CB1R activation engages β-arrestin1 to coordinate the TC-PTP-mediated inhibition of the leptin-evoked neuronal STAT3 response. This mechanism may restrict the anorexigenic effects of leptin when hypothalamic endocannabinoid levels rise, as during fasting or in diet-induced obesity.",

keywords = "Cell biology, Molecular biology",

author = "Gerg{\H o} Szanda and Tony Jourdan and {\'E}va Wisniewski and Resat Cinar and Grzegorz Godlewski and Anik{\'o} Rajki and Jie Liu and Lee Chedester and Bence Szalai and T{\'o}th, {Andr{\'a}s D{\'a}vid} and Eszter Solt{\'e}sz-Katona and L{\'a}szl{\'o} Hunyady and Asuka Inoue and Horv{\'a}th, {Vikt{\'o}ria Bea} and Andr{\'a}s Sp{\"a}t and Joseph Tam and George Kunos",

note = "Funding Information: This work was supported by the following grants: National Research, Development and Innovation Office grants NKFI-6/FK_124038 to G. Szanda, intramural NIH funds to G. Kunos and FK_ 18/128376 to {\'E}.W. (PI: Roland Cs{\'e}p{\'a}nyi-K{\"o}mi). The work was also funded by the Scientific and Innovation Fund of the Semmelweis University , Budapest, Hungary ( 26303/AOELT/2019 ; to G.S.) and the E{\"o}tv{\"o}s Lor{\'a}nd Research Network (to ELKH-SE Laboratory of Molecular Physiology Research Group ). G. Szanda was supported by the J{\'a}nos B{\'o}lyai Research Scholarship of the Hungarian Academy of Sciences . The high-content imaging system was procured using Hungarian National Competitiveness and Excellence Programme funds ( NVKP_16-1-2016-0039 ). Funding Information: We are indebted to Dr. Katalin Erd{\'e}lyi (National Institute of Oncology, Budapest, Hungary), Dr. P{\'a}l Pacher (NIAAA, NIH, Rockville, MD, USA), Dr. Bal{\'a}zs Enyedi (Semmelweis University, Budapest, Hungary) and Dr. Zolt{\'a}n Hegyi (Bio-Science, Budapest, Hungary) for technical help and valuable discussion. GT1-7 cells were kindly provided by Dr. Pamela L. Mellon (University of California, San Diego, CA, USA). The authors thank Dr. Gerhard M{\"u}ller-Newen (University Hospital Aachen, Aachen, Germany) for generously providing the STAT3-CFP-YFP construct and Dr. Tam{\'a}s Balla (NICHD, NIH, Bethesda, MD, USA) for his help in construct design and production. The CFP-tagged mouse leptin receptor clone was a generous gift from Dr. Arieh Gertler (The Hebrew University of Jerusalem, Rehovot, Israel). We are indebted to Dr. Erzs{\'e}bet Ligeti and Dr. Roland Cs{\'e}p{\'a}nyi-K{\"o}mi (Semmelweis University, Budapest, Hungary) for providing resources for molecular biological work. This work was supported by the following grants: National Research, Development and Innovation Office grants NKFI-6/FK_124038 to G. Szanda, intramural NIH funds to G. Kunos and FK_18/128376 to {\'E}.W. (PI: Roland Cs{\'e}p{\'a}nyi-K{\"o}mi). The work was also funded by the Scientific and Innovation Fund of the Semmelweis University, Budapest, Hungary (26303/AOELT/2019; to G.S.) and the E{\"o}tv{\"o}s Lor{\'a}nd Research Network (to ELKH-SE Laboratory of Molecular Physiology Research Group). G. Szanda was supported by the J{\'a}nos B{\'o}lyai Research Scholarship of the Hungarian Academy of Sciences. The high-content imaging system was procured using Hungarian National Competitiveness and Excellence Programme funds (NVKP_16-1-2016-0039). The Servier Medical Art Collection (https://smart.servier.com/) was used for the preparation of the graphical abstract. Conceptualization G.S. and G.K.; Methodology G.S. T.J. J.T. R.C. and G.K.; Formal Analysis G.S. A.S. E.S-K. A.D.T. and G.K. Investigation G.S. T.J. R.C. G.G. A.R. J.L. L.C. B.S. E.S-K. A.D.T. V.B.H. and J.T.; Resources G.S. E.W. L.H. A.I. and G.K.; Writing – Original Draft G.S. and G.K.; Writing – Review & Editing G.S. T.J. A.S. A.D.T. J.T. and G.K. Visualization G.S.; Supervision G.S. and G.K.; Funding Acquisition G.S. L.H. and G.K. All authors had access to the manuscript and agreed with the final version. The authors declare no competing interest. We support inclusive, diverse and equitable conduct of research. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = jul,

day = "21",

doi = "10.1016/j.isci.2023.107207",

language = "American English",

volume = "26",

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Szanda, G, Jourdan, T, Wisniewski, É, Cinar, R, Godlewski, G, Rajki, A, Liu, J, Chedester, L, Szalai, B, Tóth, AD, Soltész-Katona, E, Hunyady, L, Inoue, A, Horváth, VB, Spät, A, Tam, J & Kunos, G 2023, 'Cannabinoid receptor type 1 (CB₁R) inhibits hypothalamic leptin signaling via β-arrestin1 in complex with TC-PTP and STAT3', iScience, vol. 26, no. 7, 107207. https://doi.org/10.1016/j.isci.2023.107207

TY - JOUR

T1 - Cannabinoid receptor type 1 (CB1R) inhibits hypothalamic leptin signaling via β-arrestin1 in complex with TC-PTP and STAT3

AU - Szanda, Gergő

AU - Jourdan, Tony

AU - Wisniewski, Éva

AU - Cinar, Resat

AU - Godlewski, Grzegorz

AU - Rajki, Anikó

AU - Liu, Jie

AU - Chedester, Lee

AU - Szalai, Bence

AU - Tóth, András Dávid

AU - Soltész-Katona, Eszter

AU - Hunyady, László

AU - Inoue, Asuka

AU - Horváth, Viktória Bea

AU - Spät, András

AU - Tam, Joseph

AU - Kunos, George

N1 - Funding Information: This work was supported by the following grants: National Research, Development and Innovation Office grants NKFI-6/FK_124038 to G. Szanda, intramural NIH funds to G. Kunos and FK_ 18/128376 to É.W. (PI: Roland Csépányi-Kömi). The work was also funded by the Scientific and Innovation Fund of the Semmelweis University , Budapest, Hungary ( 26303/AOELT/2019 ; to G.S.) and the Eötvös Loránd Research Network (to ELKH-SE Laboratory of Molecular Physiology Research Group ). G. Szanda was supported by the János Bólyai Research Scholarship of the Hungarian Academy of Sciences . The high-content imaging system was procured using Hungarian National Competitiveness and Excellence Programme funds ( NVKP_16-1-2016-0039 ). Funding Information: We are indebted to Dr. Katalin Erdélyi (National Institute of Oncology, Budapest, Hungary), Dr. Pál Pacher (NIAAA, NIH, Rockville, MD, USA), Dr. Balázs Enyedi (Semmelweis University, Budapest, Hungary) and Dr. Zoltán Hegyi (Bio-Science, Budapest, Hungary) for technical help and valuable discussion. GT1-7 cells were kindly provided by Dr. Pamela L. Mellon (University of California, San Diego, CA, USA). The authors thank Dr. Gerhard Müller-Newen (University Hospital Aachen, Aachen, Germany) for generously providing the STAT3-CFP-YFP construct and Dr. Tamás Balla (NICHD, NIH, Bethesda, MD, USA) for his help in construct design and production. The CFP-tagged mouse leptin receptor clone was a generous gift from Dr. Arieh Gertler (The Hebrew University of Jerusalem, Rehovot, Israel). We are indebted to Dr. Erzsébet Ligeti and Dr. Roland Csépányi-Kömi (Semmelweis University, Budapest, Hungary) for providing resources for molecular biological work. This work was supported by the following grants: National Research, Development and Innovation Office grants NKFI-6/FK_124038 to G. Szanda, intramural NIH funds to G. Kunos and FK_18/128376 to É.W. (PI: Roland Csépányi-Kömi). The work was also funded by the Scientific and Innovation Fund of the Semmelweis University, Budapest, Hungary (26303/AOELT/2019; to G.S.) and the Eötvös Loránd Research Network (to ELKH-SE Laboratory of Molecular Physiology Research Group). G. Szanda was supported by the János Bólyai Research Scholarship of the Hungarian Academy of Sciences. The high-content imaging system was procured using Hungarian National Competitiveness and Excellence Programme funds (NVKP_16-1-2016-0039). The Servier Medical Art Collection (https://smart.servier.com/) was used for the preparation of the graphical abstract. Conceptualization G.S. and G.K.; Methodology G.S. T.J. J.T. R.C. and G.K.; Formal Analysis G.S. A.S. E.S-K. A.D.T. and G.K. Investigation G.S. T.J. R.C. G.G. A.R. J.L. L.C. B.S. E.S-K. A.D.T. V.B.H. and J.T.; Resources G.S. E.W. L.H. A.I. and G.K.; Writing – Original Draft G.S. and G.K.; Writing – Review & Editing G.S. T.J. A.S. A.D.T. J.T. and G.K. Visualization G.S.; Supervision G.S. and G.K.; Funding Acquisition G.S. L.H. and G.K. All authors had access to the manuscript and agreed with the final version. The authors declare no competing interest. We support inclusive, diverse and equitable conduct of research. Publisher Copyright: © 2023 The Author(s)

PY - 2023/7/21

Y1 - 2023/7/21

N2 - Molecular interactions between anorexigenic leptin and orexigenic endocannabinoids, although of great metabolic significance, are not well understood. We report here that hypothalamic STAT3 signaling in mice, initiated by physiological elevations of leptin, is diminished by agonists of the cannabinoid receptor 1 (CB1R). Measurement of STAT3 activation by semi-automated confocal microscopy in cultured neurons revealed that this CB1R-mediated inhibition requires both T cell protein tyrosine phosphatase (TC-PTP) and β-arrestin1 but is independent of changes in cAMP. Moreover, β-arrestin1 translocates to the nucleus upon CB1R activation and binds both STAT3 and TC-PTP. Consistently, CB1R activation failed to suppress leptin signaling in β-arrestin1 knockout mice in vivo, and in neural cells deficient in CB1R, β-arrestin1 or TC-PTP. Altogether, CB1R activation engages β-arrestin1 to coordinate the TC-PTP-mediated inhibition of the leptin-evoked neuronal STAT3 response. This mechanism may restrict the anorexigenic effects of leptin when hypothalamic endocannabinoid levels rise, as during fasting or in diet-induced obesity.

AB - Molecular interactions between anorexigenic leptin and orexigenic endocannabinoids, although of great metabolic significance, are not well understood. We report here that hypothalamic STAT3 signaling in mice, initiated by physiological elevations of leptin, is diminished by agonists of the cannabinoid receptor 1 (CB1R). Measurement of STAT3 activation by semi-automated confocal microscopy in cultured neurons revealed that this CB1R-mediated inhibition requires both T cell protein tyrosine phosphatase (TC-PTP) and β-arrestin1 but is independent of changes in cAMP. Moreover, β-arrestin1 translocates to the nucleus upon CB1R activation and binds both STAT3 and TC-PTP. Consistently, CB1R activation failed to suppress leptin signaling in β-arrestin1 knockout mice in vivo, and in neural cells deficient in CB1R, β-arrestin1 or TC-PTP. Altogether, CB1R activation engages β-arrestin1 to coordinate the TC-PTP-mediated inhibition of the leptin-evoked neuronal STAT3 response. This mechanism may restrict the anorexigenic effects of leptin when hypothalamic endocannabinoid levels rise, as during fasting or in diet-induced obesity.

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