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| Status |
Public on Aug 11, 2017 |
| Title |
Ras signaling inhibitors in in Adjuvant-induced Arthritis via targeting pathogenic antigen-specific Th17-type cells |
| Organism |
Rattus norvegicus |
| Experiment type |
Expression profiling by array
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| Summary |
The Ras family of GTPases play an important role in signaling nodes downstream to T cell receptor and CD28 activation, potentially lowering the threshold for TCR activation by autoantigens. Somatic mutation in NRAS or KRAS may cause a rare autoimmune disorder coupled with abnormal expansion of lymphocytes. T cells from Rheumatoid Arthritis (RA) patients show excessive activation of Ras/MEK/ERK pathway. The small molecule farnesylthiosalicylic acid (FTS) interferes with the interaction between Ras GTPases and their prenyl-binding chaperones to inhibit proper plasma membrane localization. In the present study, we tested the therapeutic and immunomodulatory effects of FTS and its derivative 5-fluoro-FTS (F-FTS) in the rat adjuvant-induced arthritis model (AIA). We show that AIA severity was significantly reduced by oral FTS and F-FTS treatment compared to vehicle control treatment. FTS was as effective as the mainstay anti-rheumatic drug methotrexate, and combining the two drugs significantly increased efficacy compared to each drug alone. We also discovered that FTS therapy inhibited both the CFA-driven in vivo induction of Th17 and IL-17/IFN-g producing double positive as well as the upregulation of serum levels of the Th17-associated cytokines IL-17A and IL-22. By gene microarray analysis of effector CD4+ T cells from CFA-immunized rats, re-stimulated in vitro with the mycobacterium tuberculosis heat-shock protein 65 (Bhsp65), we determined that FTS abrogated the Bhsp65-induced transcription of a large list of genes (e.g. Il17a/f, Il22, Ifng, Csf2, Lta, and Il1a). The functional enrichment bioinformatics analysis showed significant overlap with predefined gene sets related to inflammation, immune system processes and autoimmunity. In Conclusions, FTS and F-FTS display broad immunomodulatory effects in AIA with inhibition of the Th17-type response to a dominant arthritogenic antigen. Hence, targeting Ras signal-transduction cascade is a potential novel therapeutic approach for RA.
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| Overall design |
Eight-week-old Lewis rats will be injected at the base of the tail with 100 μl CFA prepared by suspending heat-killed M. tuberculosis (Difco) in mineral oil at 10 mg/ml. For determining the clinical outcomes, we will measure starting day +9 the ankle diameter and score all paws on a scale of 0–4 for degree of swelling, redness, and deformity of the joints. For drug therapy, we will randomly divide the rats into two groups. Rats in the first group will be treated daily (starting on day +1 after AIA induction) with oral FTS (100 mg/kg). Rats in the second group of control Rats will receive only the oral vehicle CMC control (0.5% carboxymethyl cellulose). At experiment end, the will be sacrificed and their spleens will be harvested for further testing. Single cell suspensions of spleen will be cultures with HSP65 (AG stimulation) or control media (Med group) for 72 hrs. Immediately thereafter, we will be process the cells for RNA isolation. Total RNA will be extracted and purified using the Direct-zol™ RNA MiniPrep w/ TRI-Reagent kits (Zymo Research Corp.) N= 8 samples ( 2 samples CMC Med, 2 samples CMC AG, 2 samples FTS Med, 2 samples FTS AG
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| Contributor(s) |
Zayoud M, Marcu-Malina V, Vax E, Jacob-Hirsch J, Elad-Sfadia G, Barshack I, Kloog Y, Goldstein I |
| Citation(s) |
28736556 |
| Submission date |
Jun 21, 2017 |
| Last update date |
Oct 25, 2022 |
| Contact name |
jasmine Jacob |
| E-mail(s) |
j-jacob@sheba.health.gov.il
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| Phone |
0523790500
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| Organization name |
Sheba Medical Center at Tel HaShomer
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| Street address |
haChevel 33
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| City |
Shoham |
| State/province |
-Select- |
| ZIP/Postal code |
60850 |
| Country |
Israel |
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| Platforms (1) |
| GPL17117 |
[RaGene-2_0-st] Affymetrix Rat Gene 2.0 ST Array [transcript (gene) version] |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA391245 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE100280_RAW.tar |
71.3 Mb |
(http)(custom) |
TAR (of CEL) |
| GSE100280_fold_change.txt.gz |
2.1 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
| Processed data are available on Series record |
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