If relevant, AIRE expression in thymic B cells may repress or induce the expression of other genes, as the induction of TRAs expression in medullary thymic epithelial cells (mTECs).8 Little is known ...about the nature of TRAs expressed by murine thymic B cells, and there is only a very limited overlap of TRAs between mTECs and extrathymic Aire-expressing cells,3 suggesting that thymic B cells may regulate the expression of a unique set of TRAs.\n50 4.62 x 10-21 ZMIZ1 Zinc finger, MIZ-type containing 1 5.49 9.73 x 10-54 PER1 Period circadian clock 1 5.43 2.24 x 10-10 LRP8 Low-density lipoprotein receptor-related protein 8, apolipoprotein e receptor 5.41 6.15 x 10-15 FOLR2 Folate receptor 2 (fetal) 5.38 9.01 x 10-08 TIE1 Tyrosine kinase with immunoglobulin-like and EGF-like domains 1 5.35 5.38 x 10-07 WDR62 WD repeat domain 62 5.33 2.73 x 10-09 SMIM24 Small integral membrane protein 24 5.29 2.67 x 10-05 ROR1 Receptor tyrosine kinase-like orphan receptor 1 5.11 1.08 x 10-20 INPP1 Inositol polyphosphate-1-phosphatase 5.06 3.31 x 10-18 GBP2 Guanylate binding protein 2, interferon-inducible 5.03 9.60 x 10-26 HLX H2.0-like homeobox 4.99 1.35 x 10-16 CYB5R2 Cytochrome b5 reductase 2 4.93 4.08 x 10-09 TMEM132A Transmembrane protein 132A 4.92 1.79 x 10-21 LPIN1 Lipin 1 4.80 1.00 x 10-27 CSRNP1 Cysteine-serine-rich nuclear protein 1 4.75 4.98 x 10-24 PLXNB1 Plexin B1 4.74 5.54 x 10-05 BAMBI BMP and activin membrane-bound inhibitor 4.74 3.73 x 10-08 MS4A4A Membrane-spanning 4-domains, subfamily A, member 4A 4.72 2.31 x 10-04 MMP14 Matrix metallopeptidase 14 (membrane-inserted) 4.69 6.63 x 10-04 ANK2 Ankyrin 2, neuronal 4.62 7.80 x 10-04 HERC5 HECT and RLD domain containing E3 ubiquitin protein ligase 5 4.59 2.30 x 10-27 RTN2 Reticulon 2 4.59 6.66 x 10-05 CYP2J2 Cytochrome P450, family 2, subfamily J, polypeptide 2 4.59 2.71 x 10-09 EPS8L1 EPS8-like 1 4.57 6.50 x 10-04 SCN1B Sodium channel, voltage gated, type I beta subunit 4.53 6.70 x 10-06 ZYX Zyxin 4.53 1.03 x 10-24 TUBA1C Tubulin, alpha 1c 4.52 1.14 x 10-06 SPHK1 Sphingosine kinase 1 4.49 4.47 x 10-05 STIL SCL/TAL1 interrupting locus 4.46 3.05 x 10-03 TLN2 Talin 2 4.43 5.89 x 10-05 CIT Citron rho-interacting serine/threonine kinase 4.41 5.57 x 10-03 AKAP12low *E12,E13 A kinase (PRKA) anchor protein 12 4.40 7.01 x 10-08 NAB2 NGFI-A binding protein 2 (EGR1 binding protein 2) 4.38 5.71 x 10-10 PDE9A Phosphodiesterase 9A 4.36 6.26 x 10-07 NET1 Neuroepithelial cell transforming 1 4.32 2.17 x 10-14 CISH Cytokine inducible SH2-containing protein 4.28 4.96 x 10-11 ABCG1 ATP-binding cassette, subfamily G (WHITE), member 1 4.27 2.57 x 10-30 NLRP2 NLR family, pyrin domain containing 2 4.26 8.94 x 10-08 PAK6 p21 protein (Cdc42/Rac)-activated kinase 6 4.24 4.01 x 10-06 IZUMO4 IZUMO family member 4 4.22 6.26 x 10-15 CALHM2 Calcium homeostasis modulator 2 4.20 4.17 x 10-10 ZDHHC11 Zinc finger, DHHC-type containing 11 4.20 2.63 x 10-03 LRRC36 Leucine rich repeat containing 36 4.20 1.15 x 10-16 MS4A14 Membrane-spanning 4-domains, subfamily A, member 14 4.20 3.88 x 10-13 FAM101B Family with sequence similarity 101, member B 4.16 4.65 x 10-06 ESR1 Estrogen receptor 1 4.16 7.03 x 10-08 METRNLlow *E14 Meteorin, glial cell differentiation regulator-like 4.13 1.92 x 10-08 KLF4 Kruppel-like factor 4 (gut) 4.10 2.91 x 10-02 SH2D2A SH2 domain containing 2A 4.09 1.79 x 10-04 PTP4A3 Protein tyrosine phosphatase type IVA, member 3 4.03 6.07 x 10-10 Table E2 List of TRAs expressed in thymic B cells compared with peripheral B cells
BackgroundDespite the promising efficacy of immune checkpoint blockers (ICB), tumor resistance and immune-related adverse events hinder their success in cancer treatment. To address these challenges, ...intratumoral delivery of immunotherapies has emerged as a potential solution, aiming to mitigate side effects through reduced systemic exposure while increasing effectiveness by enhancing local bioavailability. However, a comprehensive understanding of the local and systemic distribution of ICBs following intratumoral administration, as well as their impact on distant tumors, remains crucial for optimizing their therapeutic potential.To comprehensively investigate the distribution patterns following the intratumoral and intravenous administration of radiolabeled anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and to assess its corresponding efficacy in both injected and non-injected tumors, we conducted an immunoPET imaging study.MethodsCT26 and MC38 syngeneic colorectal tumor cells were implanted subcutaneously on both flanks of Balb/c and C57Bl/6 mice, respectively. Hamster anti-mouse CTLA-4 antibody (9H10) labeled with zirconium-89 (89Zr9H10) was intratumorally or intravenously administered. Whole-body distribution of the antibody was monitored by immunoPET imaging (n=12 CT26 Balb/c mice, n=10 MC38 C57Bl/6 mice). Tumorous responses to injected doses (1–10 mg/kg) were correlated with specific uptake of 89Zr9H10 (n=24). Impacts on the tumor microenvironment were assessed by immunofluorescence and flow cytometry.ResultsHalf of the dose was cleared into the blood 1 hour after intratumoral administration. Despite this, 7 days post-injection, 6–8% of the dose remained in the intratumoral-injected tumors. CT26 tumors with prolonged ICB exposure demonstrated complete responses. Seven days post-injection, the contralateral non-injected tumor uptake of the ICB was comparable to the one achieved through intravenous administration (7.5±1.7% ID.cm–3 and 7.6±2.1% ID.cm–3, respectively) at the same dose in the CT26 model. This observation was confirmed in the MC38 model. Consistent intratumoral pharmacodynamic effects were observed in both intratumoral and intravenous treatment groups, as evidenced by a notable increase in CD8+T cells within the CT26 tumors following treatment.ConclusionsImmunoPET-derived pharmacokinetics supports intratumoral injection of ICBs to decrease systemic exposure while maintaining efficacy compared with intravenous. Intratumoral-ICBs lead to high local drug exposure while maintaining significant therapeutic exposure in non-injected tumors. This immunoPET approach is applicable for clinical practice to support evidence-based drug development.
BackgroundIntratumorous immunotherapy for cancer is currently thriving. The aim of such local strategy is to improve the therapeutic index of these treatments, for higher on-target/on-tumor activity ...and less on-target/off-tumor adverse events. Strategies allowing for slow release of anti-CTLA4 in the tumor microenvironment could improve their clinical efficacy.The purpose of the study was to develop a radiopaque delivery platform to improve the targeting and exposure of intratumorous anti-CTLA4 antibodies for cancer immunotherapy.MethodsPickering emulsions of anti-CTLA4 antibodies were formulated with radiopaque ethiodized oil and poly-lactic-co-glycolic acid (PLGA) nanoparticles. We characterized the microscopic aspect and stability of such emulsions using Turbiscan. We monitored the release of anti-CTLA4 over time from these emulsions and evaluated their structure using mass spectrometry. We then tested the functionality of the released antibodies by preforming ex vivo competitive binding assays. Finally, we assessed the in vivo efficacy of intratumorous anti-CTLA4 Pickering emulsions.ResultsPickering emulsions of ethiodized oil and PLGA nanoparticles (PEEPs) resulted in a radiopaque water-in-oil emulsion with average internal phase droplet size of 42±5 µm at day 7. Confocal microscopy showed that anti-CTLA4 antibodies were effectively encapsulated by ethiodized oil with PLGA nanoparticles located at the interface between the aqueous and the oily phase. Turbiscan analysis showed that emulsions were stable with continuous and progressive release of anti-CTLA4 antibodies reaching 70% at 3 weeks. Structural and functional analysis of the released antibodies did not show significant differences with native anti-CTLA4 antibodies. Finally, intratumorous anti-CTLA4 PEEPs were able to eradicate tumors and cure mice in a syngeneic immunocompetent preclinical tumor model.ConclusionPickering emulsions of ethiodized oil and PLGA is an innovative radiopaque delivery platform that does not alter the functionality of anti-CTLA4 immune checkpoint antibodies. Beyond local anti-CTLA4 applications, these emulsions might be used with other therapeutic molecules for optimal intratumorous or intra-arterial delivery of novel cancer immunotherapies.
BackgroundPatients with high-risk non muscle-invasive bladder cancer (NMIBC) frequently relapse after standard BCG immunotherapy and have a dismal outcome after progression to muscle-invasive bladder ...cancer (MIBC).1 2 The mechanisms of tumor resistance to such immunotherapy remain elusive.MethodsWe performed functional assays of fresh human bladder tumors mixed with BCG, reinforced with in vitro experiments and in situ transcriptomics analyses together with immune profiling by immunohistochemistry (IHC) in a cohort of T1 NMIBC pre- and post BCG therapy.ResultsWe found two distinct patterns of BCG-induced immune subversion. In the first pattern, intracellular infection by live BCG was associated with HLA-I loss and epithelial-to-mesenchymal transition characteristics. Mechanistically, LC3-GFP reporter cell line showed a significant induction of autophagy upon BCG exposure. HLA-I deficient tumors displayed a myeloid immunosuppressive microenvironment together with an upregulation of autophagy-related genes, and dismal outcome. Conversely, HLA-I+ BCG-treated tumors generated a Th1 type of immune response associated with an upregulation of exhaustion markers. Such patients had a very favorable outcome upon radical surgery.ConclusionsWe surmise that HLA-I expression in bladder cancers does not result from immunoediting but rather from HLA-I molecules endocytosis related to autophagy induction in infected cancer cells. Cancer cells HLA-I scoring by immunohistochemistry staining can be easily implemented by pathologists in routine practice to stratify future bladder cancer patient treatment strategies.ReferencesPietzak EJ, Zabor EC, Bagrodia A, et al. Genomic differences between “primary” and “secondary” muscle-invasive bladder cancer as a basis for disparate outcomes to cisplatin-based neoadjuvant chemotherapy. Eur Urol 2019;75(2):231–239.. Patrick J Hensley, Kelly K Bree, Matthew T Campbell, et al. Progression of disease after BCG therapy: refining patient selection for neoadjuvant chemotherapy before radical cystectomy. J Urol 2021 June 29;101097JU0000000000001943.Ethics ApprovalOur study obtained ethics approval from the Foch Hospital Ethics Committee (IRB00012437). All the participants gave informed consent before taking part.ConsentWe surmise that HLA-I expression in bladder cancers does not result from immunoediting but rather from HLA-I molecules endocytosis related to autophagy induction in infected cancer cells. Cancer cells HLA-I scoring by immunohistochemistry staining can be easily implemented by pathologists in routine practice to stratify future bladder cancer patient treatment strategies.
Human intratumoral immunotherapy (HIT-IT) is under rapid development, with promising preliminary results and high expectations for current phase III trials. While outcomes remain paramount for ...patients and the referring oncologists, the technical aspects of drug injection are critical to the interventional radiologist to ensure optimal and reproducible outcomes. The technical considerations for HIT-IT affect the safety, efficacy, and further development of this treatment option. Image-guided access to the tumor allows the therapeutic index of a treatment to be enhanced by increasing the intratumoral drug concentration while minimizing its systemic exposure and associated on-target off-tumor adverse events. Direct access to the tumor also enables the acquisition of cancer tissue for sequential sampling to better understand the pharmacodynamics of the injected immunotherapy and its efficacy through correlation of immune responses, pathologic responses, and imaging tumor response. The aim of this article is to share the technical insights of HIT-IT, with particular consideration for patient selection, lesion assessment, image guidance, and technical injection options. In addition, the organization of a standard patient workflow is discussed, so as to optimize HIT-IT outcome and the patient experience.
Interventional radiology techniques provide excellent local tumor control for small tumors in various organs, but several limitations can hamper the oncological outcomes such as the tumor size or the ...number of lesions. Technical improvements, optimal patient selection and combination with systemic therapies, including immune checkpoint inhibitors, have been successfully developed to overcome these barriers.In this setting, chemotherapy and targeted therapies aim to diminish the tumor burden in addition to local treatments, while immunotherapies may have a synergistic effect in terms of mechanism of action on the tumor cell as well as the immune environment, with multiple treatment combinations being available. Finally, interventional Rrdiology treatments often increase tumor antigen exposure to the immune system, and thus stimulate a specific antitumor immune response that can act beyond the treated site. Notwithstanding their many benefits, combination treatment may also result in complications, the most feared may be auto-immune-related adverse events.In early studies, several combined therapies have shown promising levels of safety and efficacy, particularly in hepatocellular carcinoma.This review provides a comprehensive and up-to-date overview of results of combined therapies for primary and secondary liver malignancies. Recent advances and future perspectives will be discussed.
Many intratumoural (IT) immunotherapies are currently developed in the clinic with the aim of overcoming primary and secondary resistance and/or to limit on-target/off-tumour toxicities of immune ...checkpoint targeted therapies. This study aimed to describe the feasibility, safety and efficacy of IT immunotherapy treatments.
This retrospective single-centre study included the first 100 consecutive patients enrolled in Gustave Roussy's Human IntraTumoral-ImmunoTherapy (HIT-IT) program. Patient characteristics, target description, image guidance, safety and response according to iRECIST (Response Evaluation Criteria in Solid Tumours for immunotherapy trials) were recorded. Predictive factors of complications and responses were analysed. Survival was also reported.
From 09/2015 to 05/2020, 100 patients had 115 tumours injected during 423 treatment cycles. Most frequent primary tumour arose from the skin (n = 49), digestive track (n = 4) or head and neck (n = 8). Injected tumours’ mean diameter was 37 ± 23 mm, and a median number of 4 IT injections per patient (interquartile range:3–5) were performed. Targeted tumours for IT injections were superficial lymph nodes (36.5%), subcutaneous lesions (25.2%), liver tumours (20.9%) and others (17.4% including tumour sites such as deep lymph nodes or lung). Most patients (72%) received systemic immunotherapy in combination with HIT-IT. Procedure- and drug-related adverse events (AEs) occurred in 11.3% and 33.3% of the treatment cycles, respectively. Only 3 procedure-related AEs were grade-3 (0.7%); and no grade-4 or 5 occurred. Among all cycles, 7 grade-3 and 1 grade-5 drug-related AEs were reported. Complete and partial responses were achieved for 5% and 18% of patients, respectively, while stable disease was the best response for 11%. Patients receiving HIT-IT as a 1st-line treatment (24%), or not previously pre-treated with immunotherapy (53%) responded better, p = 0.001 and p = 0.004, respectively. From 1st cycle of IT, 12-month overall progression-free survival and overall survival were 21% (14–31%) and 57% (47–68%), respectively.
This retrospective study, conducted on patients with cancer and treated within clinical trials at Gustave Roussy, demonstrates the feasibility and safety of the IT immunotherapy strategy.
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•Image-guided intratumoural immunotherapy is feasible for both superficial and deep lesions.•No procedure-related grade 4 or 5 adverse events were observed.•A high response rate of injected tumours has been observed.
Patients with high-risk, nonmuscle-invasive bladder cancer (NMIBC) frequently relapse after standard intravesical bacillus Calmette-Guérin (BCG) therapy and may have a dismal outcome. The mechanisms ...of resistance to such immunotherapy remain poorly understood. Here, using cancer cell lines, freshly resected human bladder tumors, and samples from cohorts of patients with bladder cancer before and after BCG therapy, we demonstrate 2 distinct patterns of immune subversion upon BCG relapse. In the first pattern, intracellular BCG infection of cancer cells induced a posttranscriptional downregulation of HLA-I membrane expression via inhibition of autophagy flux. Patients with HLA-I-deficient cancer cells following BCG therapy had a myeloid immunosuppressive tumor microenvironment (TME) with epithelial-mesenchymal transition (EMT) characteristics and dismal outcomes. Conversely, patients with HLA-I-proficient cancer cells after BCG therapy presented with CD8+ T cell tumor infiltrates, upregulation of inflammatory cytokines, and immune checkpoint-inhibitory molecules. The latter patients had a very favorable outcome. We surmise that HLA-I expression in bladder cancers at relapse following BCG does not result from immunoediting but rather from an immune subversion process directly induced by BCG on cancer cells, which predicts a dismal prognosis. HLA-I scoring of cancer cells by IHC staining can be easily implemented by pathologists in routine practice to stratify future treatment strategies for patients with urothelial cancer.
Purpose of Review
This review presents the rationale for intratumoral immunotherapy, technical considerations and safety. Clinical results from the latest trials are provided and discussed.
Recent ...Findings
Intratumoral immunotherapy is feasible and safe in a wide range of cancer histologies and locations, including lung and liver. Studies mainly focused on multi-metastatic patients, with some positive trials such as T-VEC in melanoma, but evidence of clinical benefit is still lacking. Recent results showed improved outcomes in patients with a low tumor burden.
Summary
Intratumoral immunotherapy can lower systemic toxicities and boost local and systemic immune responses. Several studies have proven the feasibility, repeatability, and safety of this approach, with some promising results in clinical trials. The clinical benefit might be improved in patients with a low tumor burden. Future clinical trials should focus on adequate timing of treatment delivery during the course of the disease, particularly in the neoadjuvant setting.
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