Long-Term Expansion of Murine Primary Hepatocyte Organoids

Kluiver, Thomas A.; Kraaier, Lianne; Peng, Weng Chuan

doi:10.1007/978-1-0716-2557-6_1

Cited by 2 publications

(3 citation statements)

References 15 publications

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“…In vitro pediatric cancer models, including neuroblastoma (NBL), acute myeloid leukemia (AML), B-cell/T-cell acute lymphoblastic leukemia (B-ALL, T-ALL), diffuse midline glioma (DMG), hepatocellular carcinoma (HCC), hepatoblastoma (HBL), and malignant rhabdoid tumors (MRT), were included; origin and culture conditions are summarized in table 1. [15][16][17][18][19][20][21][22][23][24] The adult chronic myeloid leukemia K562 line and adult T-ALL Jurkat line were used as a positive control. 6 MR1 allelic variant expression of cell lines was derived from public databases (CCLE and Crown Bioscience).…”

Section: Cell Culturementioning

confidence: 99%

“…The medium consisted of the following: Advanced DMEM/F12 supplemented with 1% P/S, 1% GlutaMAX, 10 mM HEPES, 1 x B27 supplement, 1x N2 supplement (all Life Technologies), 0.2% Normocin (Invivogen), 1.25 mM N-acetyl-L-cysteine, 10% (vol/vol) RSPO1 conditioned medium, 10 mM nicotinamide, 10 nM recombinant human (Leu15)-gastrin I (All Sigma), 50 ng/mL recombinant human EGF, 25 ng/mL recombinant human HGF (Both Peprotech), 10 µM forskolin, 5 µM A8301 (Both Tocris Bioscience), 10 µM Y27632 ((Abmole), and 0.5 nM WNT surrogate. 18 Cell culture and passing were performed as described by Kluiver et al 19 . Basement membrane extract (BME) (R&D systems) 3D-cultures were dissociated using dispase and TripleE (both Life Technologies).…”

Section: Cytokine Secretion Assaymentioning

confidence: 99%

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Targeting pediatric cancers via T-cell recognition of the monomorphic MHC class I-related protein MR1

Cornel,

van der Sman,

van Dinter

et al. 2024

J Immunother Cancer

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Human leukocyte antigen (HLA) restriction of conventional T-cell targeting introduces complexity in generating T-cell therapy strategies for patients with cancer with diverse HLA-backgrounds. A subpopulation of atypical, major histocompatibility complex-I related protein 1 (MR1)-restricted T-cells, distinctive from mucosal-associated invariant T-cells (MAITs), was recently identified recognizing currently unidentified MR1-presented cancer-specific metabolites. It is hypothesized that the MC.7.G5 MR1T-clone has potential as a pan-cancer, pan-population T-cell immunotherapy approach. These cells are irresponsive to healthy tissue while conferring T-cell receptor(TCR) dependent, HLA-independent cytotoxicity to a wide range of adult cancers. Studies so far are limited to adult malignancies. Here, we investigated the potential of MR1-targeting cellular therapy strategies in pediatric cancer. Bulk RNA sequencing data of primary pediatric tumors were analyzed to assessMR1expression.In vitropediatric tumor models were subsequently screened to evaluate their susceptibility to engineered MC.7.G5 TCR-expressing T-cells. Targeting capacity was correlated with qPCR-basedMR1mRNA and protein overexpression. RNA expression ofMR1in primary pediatric tumors varied widely within and between tumor entities. Notably, embryonal tumors exhibited significantly lowerMR1expression than other pediatric tumors. In line with this, most screened embryonal tumors displayed resistance to MR1T-targetingin vitro. MR1T susceptibility was observed particularly in pediatric leukemia and diffuse midline glioma models. This study demonstrates potential of MC.7.G5 MR1T-cell immunotherapy in pediatric leukemias and diffuse midline glioma, while activity against embryonal tumors was limited. The dismal prognosis associated with relapsed/refractory leukemias and high-grade brain tumors highlights the promise to improve survival rates of children with these cancers.

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Section: Cell Culturementioning

confidence: 99%

Section: Cytokine Secretion Assaymentioning

confidence: 99%

Targeting pediatric cancers via T-cell recognition of the monomorphic MHC class I-related protein MR1

Cornel,

van der Sman,

van Dinter

et al. 2024

J Immunother Cancer

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“…Organoids could be passaged every 1-3 weeks and varied between tumors. Passaging was performed as described before 69 . Briefly, BME was first dissociated using dispase (STEMCELL Technologies), before washing at least twice with PBS supplemented with 5 % FBS (v/v).…”

Section: Primary Tumor Tissuesmentioning

confidence: 99%

Multi-dimensional profiling of hepatoblastomas and patient-derived tumor organoids uncovers tumor subpopulations with divergent WNT activation profiles and identifies pan-hepatoblastoma drug sensitivities

Kluiver,

Lu,

Schubert

et al. 2023

Preprint

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SUMMARYHepatoblastoma, the most prevalent pediatric liver cancer, almost always carries a WNT-activatingCTNNB1mutation, yet exhibits notable molecular heterogeneity. To characterize this heterogeneity and identify novel targeted therapies, we performed comprehensive analysis of hepatoblastomas and tumor-derived organoids using single-cell RNA-seq, spatial transcriptomics, single-cell ATAC-seq and high throughput drug profiling. We identified two distinct tumor epithelial signatures: hepatic ‘fetal-like’ and WNT-high ‘embryonal-like’ signatures, displaying divergent WNT signaling patterns. The liver-specific WNT targets were enriched in the fetal-like group, while the embryonal-like group was enriched in canonical WNT target genes. Gene regulatory network analysis revealed enrichment of regulons related to hepatic function such as bile acid, lipid and xenobiotic metabolism in the fetal-like subgroup but not in the embryonal-like subgroup. In addition, the dichotomous expression pattern of the transcription factors HNF4A and LEF1 allowed for a clear distinction between the fetal- and embryonal-like tumors. We also performed high-throughput drug screening using patient-derived tumor organoids and identified sensitivity to multiple inhibitor classes, most notably HDAC inhibitors. Intriguingly, embryonal-like tumor organoids, but not fetal-like tumor organoids, were sensitive to FGFR inhibitor treatments, suggesting a dependency on FGFR signaling. In summary, our data uncover the molecular and drug sensitivity landscapes of hepatoblastoma and pave the way for the development of targeted therapies.

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Long-Term Expansion of Murine Primary Hepatocyte Organoids

Cited by 2 publications

References 15 publications

Targeting pediatric cancers via T-cell recognition of the monomorphic MHC class I-related protein MR1

Targeting pediatric cancers via T-cell recognition of the monomorphic MHC class I-related protein MR1

Multi-dimensional profiling of hepatoblastomas and patient-derived tumor organoids uncovers tumor subpopulations with divergent WNT activation profiles and identifies pan-hepatoblastoma drug sensitivities

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