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Siponimod (Mayzent) Downregulates RhoA and Cell Surface Expression of the S1P1 and CX3CR1 Receptors in Mouse RAW 264.7 Macrophages

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Archivum Immunologiae et Therapiae Experimentalis Aims and scope

Abstract

The Siponimod (Mayzent) is a newly developed drug, similar to Fingolimod (FTY720) but with fewer side effects, approved by the Food and Drug Administration for the treatment of multiple sclerosis (MS). The therapeutic effect of siponimod and FTY720 in MS relies on their inhibitory effect on the sphingosine 1-phosphate (S1P) signaling. These drugs bind to the S1P receptors and block the CCL2 chemokine pathway that is responsible for the exit of the immune cells from the lymphoid organs, and circulation, thus preventing immune cell-dependent injury to the nervous system. We recently found that FTY720 beside its effect on the S1P pathway also blocks the RhoA pathway, which is involved in the actin cytoskeleton-related function of macrophages, such as expression/recycling of fractalkine (CX3CL1) receptors (CX3CR1), which direct macrophages to the transplanted organs during the development of the long-term (chronic) rejection. Here we tested the effects of siponimod on the RhoA pathway and the expression of the S1P1 and CX3CR1 receptors in mouse RAW 264.7 macrophages. We found that siponimod downregulates the expression of RhoA protein and decreases the cell surface expression of S1P1 and CX3CR1 receptors. This newly discovered crosstalk between S1P and RhoA/CX3CR1 pathways may help in the development of novel anti-chronic rejection therapies in clinical transplantation.

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Abbreviations

CCL2:

C–C motif chemokine 2

CCR2:

C–C motif chemokine 2 receptor

CX3CL1:

C–X3–C motif chemokine ligand 1, fractalkine

CX3CR1:

C–X3–C motif chemokine receptor 1, fractalkine receptor

FTY720:

Fingolimod

GAP:

GTPase-activating protein

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GEF:

Guanine nucleotide exchange factor

HRP:

Horseradish peroxidase

MS:

Multiple sclerosis

Rac1:

Rac family small GTPase 1

RAW 264.7:

Abelson murine leukemia transformed mouse macrophages

RhoA:

Ras homolog family member A

ROCK:

Rho-associated protein kinase

S1P:

Sphingosine 1-phosphate

S1P1:

Sphingosine 1-phosphate receptor 1

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Acknowledgements

We acknowledge that some of the images used to make figures were from the Servier Medical ART: SMART, http://smart.servier.com. We are grateful for the support from The William Stamps Farish Fund and we acknowledge the HMRI Flow Cytometry Core. We also acknowledge that some of the images used to make figures were from the Servier Medical ART: SMART, smart.servier.com.

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Authors and Affiliations

  1. The Houston Methodist Research Institute, Houston, TX, USA

    Ahmed Uosef, Nicole Vaughn, Xiufeng Chu, Mahmoud Elshawwaf, Ahmed Adel Abbas Abdelshafy, Kamal Mamdoh Kamal Elsaid, Rafik Mark Ghobrial & Malgorzata Kloc

  2. Department of Surgery, Houston Methodist Hospital, 6550 Fannin St., Houston, TX, 77030, USA

    Ahmed Uosef, Mahmoud Elshawwaf, Ahmed Adel Abbas Abdelshafy, Kamal Mamdoh Kamal Elsaid, Rafik Mark Ghobrial & Malgorzata Kloc

  3. Department of General Surgery, Faculty of Medicine, Ain-Shams University, Cairo, Egypt

    Ahmed Adel Abbas Abdelshafy & Kamal Mamdoh Kamal Elsaid

  4. Department of Genetics, The University of Texas, M.D. Anderson Cancer Center, Houston, TX, USA

    Malgorzata Kloc

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  1. Ahmed Uosef
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  2. Nicole Vaughn
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Correspondence to Rafik Mark Ghobrial or Malgorzata Kloc.

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Uosef, A., Vaughn, N., Chu, X. et al. Siponimod (Mayzent) Downregulates RhoA and Cell Surface Expression of the S1P1 and CX3CR1 Receptors in Mouse RAW 264.7 Macrophages. Arch. Immunol. Ther. Exp. 68, 19 (2020). https://doi.org/10.1007/s00005-020-00584-4

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