Subject(s)
Glioma , Humans , Child , Glioma/drug therapy , Glioma/genetics , Benzamides/pharmacology , Indazoles , Spinal Cord , Rho Guanine Nucleotide Exchange FactorsABSTRACT
Most cases of rhabdomyosarcoma (RMS) are sporadic and not associated with the Lynch syndrome (LS) spectrum. We report a young adult patient with RMS and a family history of colorectal cancer. Comprehensive cancer genomic profiling (CGP) of his tumor revealed a likely pathogenic variant of MSH2, NM_000251.3:c.1741delA (p.I581Lfs*9), which was also present in his blood sample. The widespread use of CGP may reveal that RMS can be a rare manifestation of LS.
ABSTRACT
Disialoganglioside (GD2)-specific chimeric antigen receptor (CAR)-T cells (GD2-CAR-T cells) have been developed and tested in early clinical trials in patients with relapsed/refractory neuroblastoma. However, the effectiveness of immunotherapy using these cells is limited, and requires improvement. Combined therapy with CAR-T cells and molecular targeted drugs could be a promising strategy to enhance the antitumor efficacy of CAR T cell immunotherapy. Here, we generated GD2-CAR-T cells through piggyBac transposon (PB)-based gene transfer (PB-GD2-CAR-T cells), and analyzed the combined effect of these cells and a MEK inhibitor in vitro and in vivo on neuroblastoma. Trametinib, a MEK inhibitor, ameliorated the killing efficacy of PB-GD2-CAR-T cells in vitro, whereas a combined treatment of the two showed superior antitumor efficacy in a murine xenograft model compared to that of PB-GD2-CAR-T cell monotherapy, regardless of the mutation status of the MAPK pathway in tumor cells. The results presented here provide new insights into the feasibility of combined treatment with CAR-T cells and MEK inhibitors in patients with neuroblastoma.
Subject(s)
Antineoplastic Agents/therapeutic use , Gangliosides/therapeutic use , Immunotherapy, Adoptive/methods , Mitogen-Activated Protein Kinase Kinases/antagonists & inhibitors , Neuroblastoma/therapy , Pyridones/therapeutic use , Pyrimidinones/therapeutic use , Receptors, Chimeric Antigen/therapeutic use , Animals , Cell Line, Tumor , Combined Modality Therapy/methods , Coumarins/therapeutic use , DNA Transposable Elements , Drug Resistance, Neoplasm , Female , Genetic Therapy/methods , Humans , Mice , Mice, SCID , Mutation , Neoplasm Recurrence, Local/therapy , Protein Kinase Inhibitors/therapeutic use , T-Lymphocytes , Xenograft Model Antitumor Assays , ras Proteins/antagonists & inhibitorsABSTRACT
The quality of chimeric antigen receptor (CAR)-T cell products, including the expression of memory and exhaustion markers, has been shown to influence their long-term functionality. The manufacturing process of CAR-T cells should be optimized to prevent early T cell exhaustion during expansion. Activation of T cells by monoclonal antibodies is a critical step for T cell expansion, which may sometimes induce excess stimulation and exhaustion of T cells. Given that piggyBac transposon (PB)-based gene transfer could circumvent the conventional pre-activation of T cells, we established a manufacturing method of PB-mediated HER2-specific CAR-T cells (PB-HER2-CAR-T cells) that maintains their memory phenotype without early T cell exhaustion. Through stimulation of CAR-transduced T cells with autologous peripheral blood mononuclear cell-derived feeder cells expressing both truncated HER2, CD80, and 4-1BBL proteins, we could effectively propagate memory-rich, PD-1-negative PB-HER2-CAR-T cells. PB-HER2-CAR-T cells demonstrated sustained antitumor efficacy in vitro and debulked the HER2-positive tumors in vivo. Mice treated with PB-HER2-CAR-T cells rejected the second tumor establishment owing to the in vivo expansion of PB-HER2-CAR-T cells. Our simple and effective manufacturing process using PB system and genetically modified donor-derived feeder cells is a promising strategy for the use of PB-CAR-T cell therapy.
ABSTRACT
Ephrin type-B receptor 4 (EPHB4), expressed in tumors including rhabdomyosarcoma, is a suitable target for chimeric antigen receptor (CAR)-T cells. Ligand-independent activation of EPHB4 causes cell proliferation and malignant transformation in rhabdomyosarcoma, whereas ligand-dependent stimulation of EPHB4 induces apoptosis in rhabdomyosarcoma. Therefore, we hypothesized that ligand-based, EPHB4-specific CAR-T cells may kill rhabdomyosarcoma cells without stimulating downstream cell proliferation mechanisms. We developed novel CAR-T cells by targeting EPHB4 via EPHRIN B2, a natural ligand of EPHB4. The generation of EPHB4-CAR-T cells via piggyBac (PB) transposon-based gene transfer resulted in sufficient T cell expansion and CAR positivity (78.5% ± 5.9%). PB-EPHB4-CAR-T cells displayed a dominant stem cell memory fraction (59.4% ± 7.2%) as well as low PD-1 expression (0.60% ± 0.21%) after 14 days of expansion. The PB-EPHB4-CAR-T cells inhibited EPHB4-positive tumor cells without activating cell proliferation downstream of EPHB4, even after multiple tumor re-challenges and suppressed tumor growth in xenograft-bearing mice. Therefore, PB-EPHB4-CAR-T cells possess a memory-rich fraction without early T cell exhaustion and show potential as promising therapeutic agents for treating rhabdomyosarcoma and other EPHB4-positive tumors.
ABSTRACT
A 14-year-old girl was referred to our department because of headache and visual impairment following the resection of recurrent cardiac myxoma. Head magnetic resonance imaging (MRI) scan detected an intra- and supra-sellar tumor. Moreover, the patient showed the presence of spotty skin pigmentations on her cheeks and lower lip. Blood examination revealed hypothyrotropinemia, and ultrasonography results revealed multiple thyroid nodules. She was diagnosed with Carney complex (CNC). Her pituitary tumor was suspected as growth hormone (GH)-secreting adenoma, because overgrowth was observed in the patient. However, biochemical examinations, including oral glucose tolerance test, failed to show the characteristic findings of GH-secreting adenoma. In contrast, insulin tolerance test showed GH deficiency. Her visual impairment improved without performing decompression surgery, and the tumor size decreased, as per the MRI findings. Based on clinical course, the patient was diagnosed with pituitary apoplexy in pituitary adenoma, following which she was discharged. At 3 months after discharge, thyrotropin-releasing hormone loading test performed revealed low thyrotropin-stimulating hormone and thyroid hormone levels, and the patient was in a depressed mood. Therefore, l-T4 replacement was initiated, following which her GH secretory capacity gradually improved. Here, we report, to the best of our knowledge, the first case of a patient with pituitary apoplexy in CNC. Such condition must be identified in young patients with recurrent cardiac myxoma, and examinations, such as head MRI, must be performed.
Subject(s)
Cardiac Surgical Procedures/adverse effects , Carney Complex/complications , Heart Neoplasms/surgery , Myxoma/surgery , Pituitary Apoplexy/etiology , Pituitary Neoplasms/complications , Adolescent , Carney Complex/diagnosis , Female , Growth Hormone-Secreting Pituitary Adenoma/complications , Humans , Neoplasm Recurrence, Local/surgery , Pituitary Neoplasms/diagnosis , Pituitary Neoplasms/surgery , Thyrotropin/deficiencyABSTRACT
Lipoblastoma is a rare benign adipose tissue tumor that occurs mostly in infants and children. Histological diagnosis of lipoblastoma is sometimes difficult because it closely resembles other lipomatous tumors. The detection of PLAG1 gene rearrangement is useful for the diagnosis of lipoblastoma. Four PLAG1 fusion partner genes are known in lipoblastoma: HAS2 at 8q24.1, COL1A2 at 7q22, COL3A1 at 2q32, and RAB2A at 8q12. Herein, we describe a novel fusion gene in a case of lipoblastoma of left back origin. We identified a potential PLAG1 fusion partner using 5' rapid amplification of cDNA ends, and sequence analysis revealed the novel fusion gene, BOC-PLAG1. The BOC-PLAG1 fusion transcript consists of the first exon of the BOC gene fused to exon 2 or exon 3 of the PLAG1 gene. PLAG1 expression was found to be 35.7⯱â¯2.1 times higher in the tumor specimen than in human adipocytes by qRT-PCR. As a result of the translocation, the constitutively active promoter of BOC leads to PLAG1 overexpression. The identification of the BOC-PLAG1 fusion gene will lead to more accurate diagnosis of lipoblastoma.