Sema3B Expression Is Reduced in Rheumatoid Arthritis Patients and Has a Protective Role in a Murine Model of Arthritis

Ana Igea ¹, Tiago Carvalheiro ², Beatriz Malvar Fernandez ², Angela Rodriguez-Trillo ³, Trudy McGarry ⁴, Carmen Conde ⁵, Douglas Veale ⁶, Ursula Fearon ⁷, Africa Gonzalez ¹, Timothy R.D. Radstake ⁸, Kris A. Reedquist ² and Samuel Garcia⁹, ¹Department of Immunology, Centro de Investigaciones Biomédicas (CINBIO), Centro de Investigación Singular de Galicia, Instituto de Investigación Sanitaria Galicia Sur, Universidad de Vigo, Vigo, Spain, ²Department of Rheumatology and Clinical Immunology and Laboratory of Translational Immunology, University Medical Center Utrecht, University of Utrecht, Utrecht, Netherlands, ³laboratorio de Reumatología Experimental y Observacional, Instituto de Investigación Sanitaria de Santiago (IDIS)-Hospital Clinico (CHUS), Santiago de Compostela, Spain, ⁴Department of Molecular Rheumatology, Trinity Biomedical Science Institute, Trinity College Dublin, Dublin, Ireland, ⁵Laboratorio de Reumatología Experimental y Observacional, y Servicio de Reumatología, Instituto de Investigacion Sanitaria de Santiago (IDIS), Hospital Clínico Universitario de Santiago de Compostela (CHUS), SERGAS, Santiago de Compostela, Spain, ⁶EULAR Centre For Arthritis And Rheumatic Diseases and The Conway Institute, Dublin, Ireland, ⁷Molecular Rheumatology, Trinity Biomedical Sciences Institute, TCD, Dublin, Ireland, ⁸Department of Rheumatology/Clinical Immunology, University Medical Center Utrecht, Utrecht, The Netherlands., Utrecht, Netherlands, ⁹Department of Rheumatology and Clinical Immunology, University Medical Center Utrecht, University of Utrecht, Utrecht, Spain

Meeting: 2019 ACR/ARP Annual Meeting

Keywords: Animal models, rheumatoid arthritis, Rheumatoid arthritis (RA), therapeutic targeting, treatment and biomarkers

Session Information

Date: Monday, November 11, 2019

Title: RA – Animal Models Poster

Session Type: Poster Session (Monday)

Session Time: 9:00AM-11:00AM

Background/Purpose: The semaphorin family is a large group of proteins initially described in axon guidance. However, semaphorins also play a role in other processes involved in rheumatoid arthritis (RA) such as the regulation of immunity, angiogenesis, apoptosis and cell migration and invasion. Previously we demonstrated that semaphorin 3B (Sema3B) expression is reduced in the synovial tissue and synovial fluid of RA compared to undifferentiated arthritis patients and reduces the invasive capacity of RA fibroblast like synoviocytes (FLS). Here, we validated initial findings in an independent cohort of patients and determined the role of Sema3B in an in vivo model of arthritis

Methods: Sema3B expression in healthy control (HC, n=10), RA (n=10) and arthralgia patient (n=8) synovial tissue and serum was determined by qPCR and ELISA. Arthritis was induced in wild type (WT) and Sema3B-deficient mice (Sema3B KO) mice (n=10 for both) by intraperitoneal injection of 100 ml of K/BxN serum on day 0 and day 2. Mice were sacrificed on day 9 after serum transfer. mRNA expression in total joints and murine fibroblast-like synociocytes (mFLS) was determined by qPCR. mFLS proliferation and migration were determined using BrdU and wound closure assays, respectively.

Results: mRNA levels of Sema3B were significantly lower in the synovial tissue of RA patients compared to arthralgia patient (p=0.0205). Importantly, serum levels of Sema3B were also reduced in RA patients compared to HC and arthralgia patients (p=0.034 and p=0.0241, respectively) The clinical severity of serum-induced arthritis was significantly higher in Sema3B KO mice compared to WT mice (p=0.0015). This was associated with a higher joint and mFLS expression of the inflammatory mediators IL-1β, TNF, IL-6, CXCL-5 and CXCL-10 and the matrix metalloproteinases MMP-2 and MMP-3 (p-values ranging between p< 0.05 and p< 0.001). Functional experiments demonstrated a significantly higher proliferation and migration capacity (p=0.034 and p=0.0313, respectively) in the mFLS from Sema3B KO compared to WT mice. Importantly, the administration of recombinant mouse Sema3B abrogated the enhanced migratory capacity of Sema3B KO mFLS (p=0.044).

Conclusion: Our data confirm our previous results demonstrating that Sema3B expression is reduced early in RA onset and provide evidence that Sema3B has a protective role in a mouse model of arthritis. Therefore, Sema3B administration could be a novel therapeutic strategy for the treatment of RA.

A-B. Sema3B mRNA and protein levels in the synovial tissue -A- and serum -B- of RA and arthralgia patients. Data is presented as a scatter plot, where each point represents an individual patient. * p<0.05.

-A- Daily global arthritic scores of WT and Sema3B deficient -KO- mice. -B- mRNA expression of inflammatory mediators in the forepaws of control and arthritic WT or Sema3B KO mice. Data is presented as a scatter plot, where each point represents an individual mouse. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. # p < 0.05, ## p < 0.01, ### p < 0.001 and #### p < 0.0001 compared to WT control mice.

A-B. Proliferation -A- and migration -B- of mFLS from WT and Sema3B-deficient -KO- mice in the presence or absence of 10% FBS for 24 h. Data is shown as BrdU incorporation -A- or migration index -B- and presented as a scatter plot, where each point represents an individual mouse. C. Migration of mFLS stimulated with Sema3B -100 ng/ml- in the presence or absence of 10% FBS for24 h. Data is shown as migration index and presented as connected dots. * p < 0.05 compared to medium 1% FBS.

Disclosure: A. Igea, None; T. Carvalheiro, None; B. Malvar Fernandez, None; A. Rodriguez-Trillo, None; T. McGarry, None; C. Conde, None; D. Veale, Health Beacon, 1; U. Fearon, None; A. Gonzalez, None; T. Radstake, None; K. Reedquist, None; S. Garcia, None.

To cite this abstract in AMA style:

Igea A, Carvalheiro T, Malvar Fernandez B, Rodriguez-Trillo A, McGarry T, Conde C, Veale D, Fearon U, Gonzalez A, Radstake T, Reedquist K, Garcia S. Sema3B Expression Is Reduced in Rheumatoid Arthritis Patients and Has a Protective Role in a Murine Model of Arthritis [abstract]. Arthritis Rheumatol. 2019; 71 (suppl 10). https://acrabstracts.org/abstract/sema3b-expression-is-reduced-in-rheumatoid-arthritis-patients-and-has-a-protective-role-in-a-murine-model-of-arthritis/. Accessed .

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