- Letter to the Editor
- Open Access
IL-7 and CCL19-secreting CAR-T cell therapy for tumors with positive glypican-3 or mesothelin
Journal of Hematology & Oncology volume 14, Article number: 118 (2021)
Although chimeric antigen receptor (CAR)-engineered T cells have shown great success in the treatment of B cell malignancies, this strategy has limited efficacy in patients with solid tumors. In mouse CAR-T cells, IL-7 and CCL19 expression have been demonstrated to improve T cell infiltration and CAR-T cell survival in mouse tumors. Therefore, in the current study, we engineered human CAR-T cells to secrete human IL-7 and CCL19 (7 × 19) and found that these 7 × 19 CAR-T cells showed enhanced capacities of expansion and migration in vitro. Furthermore, 7 × 19 CAR-T cells showed superior tumor suppression ability compared to conventional CAR-T cells in xenografts of hepatocellular carcinoma (HCC) cell lines, primary HCC tissue samples and pancreatic carcinoma (PC) cell lines. We then initiated a phase 1 clinical trial in advanced HCC/PC/ovarian carcinoma (OC) patients with glypican-3 (GPC3) or mesothelin (MSLN) expression. In a patient with advanced HCC, anti-GPC3-7 × 19 CAR-T treatment resulted in complete tumor disappearance 30 days post intratumor injection. In a patient with advanced PC, anti-MSLN-7 × 19 CAR-T treatment resulted in almost complete tumor disappearance 240 days post-intravenous infusion. Our results demonstrated that the incorporation of 7 × 19 into CAR-T cells significantly enhanced the antitumor activity against human solid tumor. Trial registration: NCT03198546. Registered 26 June 2017, https://clinicaltrials.gov/ct2/show/NCT03198546?term=NCT03198546&draw=2&rank=1
To the editor:
A recent study showed that IL-7 and CCL19 (7 × 19) secretion promotes the infiltration and survival of murine CAR-T cells in vivo [1,2,3]. To further verify the effects of 7 × 19 secretion on human CAR-T cells, we constructed three CAR vectors targeting GPC3, MSLN, and CD20 (Fig. 1a) [4,5,6], which all contained intracellular domains of CD28 and TLR2 [7,8,9]. As the cargo capacity of the lentiviral vector is limited, human IL-7 and CCL19 linked through the 2A peptide were inserted into a separate lentiviral vector (Fig. 1a). We then cotransduced T cells with CAR and 7 × 19 lentiviruses (Fig. 1b). IL-7 and CCL19 were detected in the supernatant of 7 × 19 CAR-T cell cultures (Fig. 1c). 7 × 19 augmented the chemotaxis and expansion of CAR-T cells (Fig. 1d, 1e), but did not influence the cytotoxicity and cytokine secretion of CAR-T cells (Additional file 1: Fig. S1a, S1b, S1c).
We next adopted a GPC3+ HepG2 cell line-derived xenograft (CDX) mouse model to assess the antitumor efficacy of anti-GPC3-7 × 19 CAR-T cells. Anti-GPC3-7 × 19 CAR-T cells significantly suppressed HepG2 growth than anti-GPC3 CAR-T cells (Fig. 1f, 1g). In addition, the percentage of T cells in peripheral blood of anti-GPC3-7 × 19 group were higher than those in the anti-GPC3 group (Fig. 1h, Additional file 1: Fig. S2a). There were more infiltrated T cells in the tumors of the anti-GPC3-7 × 19 group, compared that in the anti-GPC3 group (Additional file 1: Fig. S2b). Similarly, anti-GPC3-7 × 19 CAR-T cells displayed superior antitumor efficacy in a HCC patient-derived xenograft (PDX) mouse model, compared to anti-GPC3 CAR-T cells (Additional file 1: Fig. S3). Moreover, anti-MSLN-7 × 19 CAR-T cells also displayed more potent antitumor capacity than anti-MSLN CAR-T cells in a MSLN+ AsPC-1 CDX mouse model (Fig. 1i, 1j, 1k, Additional file 1: Fig. S4).
Based on the preclinical results, we conducted a phase I clinical trial in advanced hepatocellular carcinoma (HCC), pancreatic carcinoma (PC) and ovarian carcinoma (OC) patients with GPC3 or MSLN expression to further explore the clinical safety and feasibility of 7 × 19 CAR-T cell therapy. The protocol of this trial is described in the supplemental materials and is depicted schematically (Additional file 1: Fig. S5, S6). Patients were enrolled based on the patient inclusion and exclusion criteria (Additional file 1: Table S1). Six patients were administered anti-GPC3/anti-MSLN-7 × 19 CAR-T cells (Additional file 1: Table S2).
Subject GD-G/M-001 is an advanced HCC patient with GPC3 expression (Additional file 1: Fig. S7a, S7b). Three metastatic lesions were chosen to perform computed tomography (CT) guided intratumor injection of CAR-T cells. Though the sizes of two lung nodules did not change significantly 60 days after CAR-T cells injection (Fig. 2a), the liver tumor lesion (1.2 × 1.3 cm) shrunk significantly on day 10 and completely disappeared on day 32 post anti-GPC3-7 × 19 CAR-T cells injection (Fig. 2b). The patient did not suffer any toxic effects and had staging evaluation revealing partial response (PR) according to standard Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 on CT.
Subject GD-G/M-005 is an advanced PC patient with MSLN expression (Additional file 1: Fig. S7f). This subject progressed with a local lymph node metastasis (24 × 33 mm) (Fig. 2c). Anti-MSLN-7 × 19 CAR-T cells were first infused through hepatic artery with heavy fever on the night without cytokine release syndrome (CRS) or neurotoxicity . Subsequently, he received intravenous infusions of the anti-MSLN-7 × 19 CAR-T cells every 1–2 months. After 5 times of the CAR-T cells infusions, CT staging revealed complete response (CR) on day 240 with the lymph node measured 8.3 × 9.6 mm and no other enlarged lymph nodes visible (Fig. 2c). He remained in normal condition.
The clinical course of the four other infused patients is shown in Additional file 1: Fig. S8 and described in Additional file 1: Table S3 and the Materials and Methods. Our current six-case cohort preliminary clinical study revealed no grade 2–4 adverse events or major complications; one PC patient (1/6, 16.7%) achieved CR; one HCC patient (1/6, 16.7%) achieved PR; and 2 (2/6, 33.3%) achieved steady disease (SD), demonstrating the huge therapeutic potential of 7 × 19 CAR-T cell therapy for advanced solid tumors with GPC3/MSLN expression. Due to the limited number of cases enrolled in this study, we admit this was not a comprehensive clinical trial. However, based on the safety and efficacy of 7 × 19 CAR-T cells shown by these results, we are preparing to conduct a formal phase I clinical trials of 7 × 19 CAR-T cells that specifically target liver cancer or pancreatic cancer.
Availability of data and materials
All data are available in the manuscript or the supplementary materials.
Chimeric antigen receptor
- human IL-7 and CCL19:
7 × 19
Response evaluation criteria in solid tumors
Cytokine release syndrome
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This research was supported by the National Natural Science Foundation of China (No. 81672276 to Z. Z.; No. 81961128003 and 81972672 to P. L.; No. 81872613 and 81573142 to L. Y.; No. 81872069 to H. Y.),Guangzhou Science and Technology program (No. 201803010038 to Z. Z.); Strategic Priority Research Program of the Chinese Academy of Sciences (No. XDB19030205 and XDA12050305 to P. L.); The National Major Scientific and Technological Special Project for “Significant New Drugs Development (No. 2018ZX090201002-005); Guangdong provincial Significant New Drugs Development (No. 2019B020202003 to P. L.); Guangdong Special Support Program (No. 2017TX04R102 to P. L.); Guangzhou Science and Technology Plan Project (No. 201907010042 and 2020B1212060052 to P. L.); Frontier Research Program of Guangzhou Regenerative Medicine and Health Guangdong Laboratory (No. 2018GZR110105003 to P. L.); Science and Technology Planning Project of Guangdong province, China (No. 2017B030314056 to P. L.); a grant from the University Grants Committee / Research Grants Council of the Hong Kong Special Administrative Region, China (No. AoE/M-401/20 to P. L.).
Ethics approval and consent to participate
This study was approved by the Human Ethics Committees of the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China. The patient gave their written informed consent in accordance with the Declaration of Helsinki. This study is registered at www.ClinicalTrials.gov as NCT03198546. All animal experimental protocols were performed in accordance with instruction guidelines from the China Council on Animal Care and approved by the guidelines of the Ethics Committee of Animal Experiments at Guangzhou Institutes of Biomedicine and Health.
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Pang, N., Shi, J., Qin, L. et al. IL-7 and CCL19-secreting CAR-T cell therapy for tumors with positive glypican-3 or mesothelin. J Hematol Oncol 14, 118 (2021). https://doi.org/10.1186/s13045-021-01128-9
- CAR-T cell
- Pancreatic carcinoma
- Hepatocellular carcinoma
- Complete response