Biology subjects

Kruijff, S.

Publications and source records attributed to Kruijff, S..

4 recordsLinked to original sources

bioRxiv2026

Radiation-induced interferon-I response impairs thyroid organoid function

Background and AimRadiotherapy is a standard cancer treatment, but radiation exposure to surrounding healthy tissues may lead to adverse side effects that compromise patient quality of life. In patients with head and neck cancer treated with radiotherapy, thyroid damage is a frequent complication, resulting in hypothyroidism or secondary thyroid malignancies. Although clinically recognized, the molecular mechanisms underlying these side effects remain mostly unexplored. This study aims to characterize the radiation-induced molecular alterations in thyroid organoids. MethodsBulk RNA sequencing was performed to investigate transcriptomic changes in tissue-derived thyroid organoids following gamma-irradiation. Observed changes were further validated and explored using qPCRs, western blotting, immunofluorescence, caspase 3/7 activity and organoid forming efficiency. ResultsOur findings identify interferon-{beta} (IFN-{beta}) signaling as a key mediator of radiation-induced inflammation in the thyroid. Additionally, the intrinsic apoptotic pathway was found to be the predominant mechanism of radiation-induced thyroid cell death. Notably, while IFN-{beta} exhibited a protective effect against apoptosis, it concurrently reduced thyroid stem progenitor cell potential. ConclusionsThese results highlight the dual role of IFN-{beta} signaling in modulating thyroid cell fate after irradiation, potentially promoting survival upon injury at the expense of regenerative potential.

cell biology↗

bioRxiv2025

Role of Klhl14 in senescence and epithelial-to-mesenchymal transition via TGF- modulation

KLHL14, a component of an E3-ubiquitin ligase complex, has emerged as a context-dependent oncogene or tumor suppressor, particularly important for thyroid development. Yet its role in thyroid biology remains largely unexplored. In this study, we uncover a central function for KLHL14 in maintaining thyroid epithelial identity and regulating tissue homeostasis. Using a thyroid organoid model, we show that KLHL14 is essential for the proper growth and maturation of thyroid cells. Reduction of KLHL14 expression disrupts organoid development and triggers a dual cellular response involving features of both senescence and epithelial-to-mesenchymal transition. These phenotypic changes are accompanied by increased cellular plasticity and migratory capacity. Mechanistically, we identify TGF-{beta} signaling as a key pathway activated upon KLHL14 depletion, contributing to the observed cellular reprogramming. Inhibiting TGF-{beta} restores growth and reduces markers of senescence and EMT, positioning KLHL14 as an upstream modulator of this signaling axis. These findings reveal a previously unrecognized role for KLHL14, suggesting that its dysfunction may contribute to disease progression in aggressive thyroid cancers. This work broadens our understanding of thyroid epithelial biology and provides molecular insights extendable to other tissues, highlighting KLHL14 as a potential target for therapeutic interventions in malignancies displaying the herein explored features.

cell biology↗

bioRxiv2023

Patient-Derived Medullary Thyroid Cancer Organoids; a Model for Patient-tailored Drug and PET-Tracer Screening

BackgroundMedullary thyroid carcinoma (MTC) is a neuroendocrine tumor derived from the parafollicular C-cells of the thyroid gland. PET imaging, with various PET tracers, is performed when distant metastatic disease is suspected. After the recognition of progressive disease on imaging, targeted therapy may be initiated to prolong survival. Mutations in the gene encoding the REarranged during Transfection (RET) tyrosine kinase play a key role in the development of MTC. It seems that tyrosine kinase inhibitors (TKIs) inhibit tumor proliferation, but it remains challenging to determine the best patient specific treatment option. Here, we aim to set up an in vitro MTC organoid model to study its potential for patient-tailored drug-screening and uptake of PET tracers. MethodsDispersed cells obtained from surgical MTC biopsies were suspended in Matrigel with defined medium allowing MTC organoid formation. To study putative MTC stem cells, the self-renewal potential of organoids was tested by dissociation to single cells and re-plating. To check MTC origin, MTC-specific gene expression and proteins were characterized by qPCR and immunofluorescent (IF) staining. To investigate cytotoxicity, MTC-organoids (MTOs) were exposed to various TKIs after which hormone (calcitonin and CEA) excretion levels were determined. Lastly, we evaluated cell-specific uptake of clinically used Positron Emission Tomography (PET) tracers. ResultsNine MTC biopsies were processed and cultured as MTOs. Eight MTO lines were used to determine organoid formation efficiency (OFE), which yielded a maximum OFE of 6.3% in passage 1 (p1), 5.9% in p2, and 9.4% in p3, indicating the presence of putative stem cells. IF staining showed expression of MTC-specific markers in both tissue and MTOs showing tissue resemblance. Tumor marker measurements in MTO medium showed MTC-specific production of calcitonin and CEA with changed concentrations after exposure to TKIs. Exposure to PET tracers showed significant uptake in the MTOs. ConclusionMTC organoids can be successfully cultured and resemble the tissue of origin in gene expression, protein expression and functionality. In addition, MTOs can take up PET tracers, and have the potential to be used as a prediction model for TKI treatment in the future.

cancer biology↗

bioRxiv2022

Patient-derived parathyroid organoids as tracer and drug-screening application model

Parathyroid diseases are characterized by dysregulation of calcium homeostasis and alterations in parathyroid hormone (PTH) excretion. The understanding of parathyroid hyperplastic growth and the development of parathyroid-targeted treatment and imaging tracers could benefit from in vitro models. Therefore, we aim to establish stem cell-derived, three-dimensional organoids representing human parathyroid tissue in vitro. Patient-derived hyperplastic parathyroid tissue was dispersed and parathyroid organoids (PTO) were cultured and characterized. PTO-derived cells were shown to exhibit in vitro self-renewal over several passages, indicative of the presence of putative stem cells. Immunofluorescence and RNA-sequencing confirm that PTO phenocopy hyperplastic parathyroid tissue. Exposure of PTO to increasing calcium concentrations and to PTH-lowering drugs resulted in a significantly reduced PTH excretion. Next to this, the PTO showed specific binding of 11C-methionine to the targeted receptor. Additionally, when organoids were incubated with 99mTc-sestamibi, we observed a higher uptake in PTOs from patients with a 99mTc-sestamibi positive scan compared to patients with a negative scan. These data show functionality of PTOs resembling the parathyroid. In conclusion, we present a patient-derived PTO culture, that recapitulates the originating tissue on gene and protein expression and functionality. This PTO model paves the way for future physiology studies and therapeutic target and tracer discovery.

cell biology↗