Survivin Blockade Sensitizes Rhabdomyosarcoma Cells for Lysis by Fetal Acetylcholine Receptor–Redirected T Cells

Abstract

Cellular immunotherapy may provide a strategy to overcome the poor prognosis of metastatic and recurrent rhabdomyosarcoma (RMS) under the current regimen of polychemotherapy. Because little is known about resistance mechanisms of RMS to cytotoxic T cells, we investigated RMS cell lines and biopsy specimens for expression and function of immune costimulatory receptors and anti-apoptotic molecules by RT-PCR, Western blot analysis, IHC, and cytotoxicity assays using siRNA or transfection-modified RMS cell lines, together with engineered RMS-directed cytotoxic T cells specific for the fetal acetylcholine receptor. We found that costimulatory CD80 and CD86 were consistently absent from all RMSs tested, whereas inducible T-cell co-stimulator ligand (ICOS-L; alias B7H2) was expressed by a subset of RMSs and was inducible by tumor necrosis factor a in two of five RMS cell lines. Anti-apoptotic survivin, along with other inhibitor of apoptosis (IAP) family members (cIAP1, cIAP2, and X-linked inhibitor of apoptosis protein), was overexpressed by RMS cell lines and biopsy specimens. Down-regulation of survivin by siRNA or pharmacologically in RMS cells increased their susceptibility toward a T-cell attack, whereas induction of ICOS-L did not. Treatment of RMS-bearing Rag À/À mice with fetal acetylcholine receptorespecific chimeric T cells delayed xenograft growth; however, this happened without definitive tumor eradication. Combined blockade of survivin and application of chimeric T cells in vivo suppressed tumor proliferation during survivin inhibition. In conclusion, survivin blockade provides a strategy to sensitize RMS cells for T-celle based therapy. (Am J Pathol 2013, 182: 2121e2131; http://dx

Arctigenin Enhances Chemosensitivity to Cisplatin in Human Nonsmall Lung Cancer H460 Cells through Downregulation of Survivin Expression

“…Overexpression of survivin leads to uncontrolled cancer cell growth and drug resistance . Efficient inhibition of survivin expression and its functions renders tumor cells more sensitive to anticancer drugs, leading to cell apoptosis . Recent studies showed that survivin is overexpressed and is associated with chemoresistance in NSCLC .…”

cFLIP overexpression in T cells in thymoma‐associated myasthenia gravis

“…RNA isolation from unfixed thymoma tissue, thymocytes derived thereof, PBMCs, T-cell subsets, and B cells; cDNA synthesis; qRT-PCR (relative quantification was calculated using the ΔΔCt method with Glycerinaldehyd-3-phosphat-Dehydrogenase (GAPDH) standard: TaqMan; FAST SYBR Green; Applied Biosystems, Darmstadt, Germany), and western blot were performed as described. 13 Cytosolic and nuclear proteins were isolated using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermoscientific, Karlsruhe, Germany). Mouse anti-cFLIP (G11 D16A8; Cell Signaling, Francfort, Germany), rabbit anti-p65 (C-20, sc-372; Abcam, UK ) , rabbit anti-ß-actin (New England Biolabs, Frankfurt, Germany), and PARP (poly (ADP ribose) polymerase (H-250) Santa Cruz, Heidelberg, Germany) were used for western blot.…”

“…Survival of PBMCs was measured by MTT assay (Sigma Aldrich, Munich Germany) 13 For apoptosis induction, 10 5 PBMCs from thymoma patients and blood donors were treated with 5–10 ng/mL FASLG (Sigma Aldrich, Germany) for 24 h, followed by MTT assay. FAS expression levels on PBMCs were quantified by flow cytometry, using FITC-conjugated mouse anti-human-CD95 (clone DX2; BD Pharmingen, Heidelberg Germany).…”

Targeted therapy for soft tissue sarcomas in adolescents and young adults

“…One Phase I trial that included mostly patients with bone sarcomas, but one patient with DSRCT, showed tolerability of HER2 CAR T cells 183. In solid tumor CAR T cell therapy, the tumor microenvironment seems to interfere with proper T cell function184 and this might also apply to sarcomas 179. New approaches incorporating immune-modulatory strategies, such as PD-1 or gamma delta T cells into CAR T cell therapeutic approaches,185–187 are being investigated.…”