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Rübe, C.

Publications and source records attributed to Rübe, C..

3 recordsLinked to original sources

Histone Variant H2A.J Links Epigenetic Reprogramming to Mitochondrial-dependent Kidney Regeneration under Radiation Stress

The histone variant H2A.J is implicated in radiation-induced senescence by promoting the transcription of inflammatory genes. While H2A.J expression increases in renal tubular epithelial cells (TECs) following ionizing radiation (IR), its functional role remains poorly understood. To investigate this, constitutive H2A.J knock-out (KO) mice and wild-type (WT) controls were subjected to CT-guided IR (fractionated whole-body or localized kidney IR). Kidneys were analyzed at acute, intermediate, and chronic stages using immunofluorescence, histochemistry, automated image analysis, and electron microscopy. In WT TECs, IR induced rapid chromatin incorporation and C-terminal serine phosphorylation of H2A.J. Conversely, KO kidneys exhibited significantly more severe histopathological damage, including tubular dilation, flattened epithelium, associated with increased apoptosis, and premature senescence, characterized by persistent DNA damage with lamin B1 loss. Notably, KO TECs displayed disrupted mitochondrial networks and reduced brush borders even at baseline, which were further exacerbated by IR. Unlike WT controls, KO kidneys developed progressive tubular atrophy and incipient fibrosis, indicating a failure in regenerative capacity. Our findings suggest that H2A.J loss impairs tubular regeneration due to defective mitochondrial activation, resulting in insufficient energy supply for coordinated repair. Collectively, these results identify H2A.J as a critical stress-adaptive histone variant essential for the epigenetic regulation of tissue repair following radiation-induced damage. One Sentence SummaryIn irradiated kidney, the loss of histone variant H2A.J impairs the chromatin-mediated adaptation of mitochondrial function in tubular epithelial cells, thereby exacerbating cellular stress - characterized by increased induction of apoptosis and senescence - and ultimately leading to tubular atrophy. Translational RelevanceAcute and chronic kidney injury are frequent complications of genotoxic cancer therapies. Chemo- and radiotherapy induce DNA lesions that trigger cell death and senescence, often leading to irreversible renal damage. However, renal regeneration can occur through the dedifferentiation, proliferation, and redifferentiation of surviving tubular epithelial cells (TECs). This repair process is governed by epigenetic mechanisms that regulate the DNA damage response (DDR) and adapt gene expression programs. Following ionizing radiation (IR), epigenetic remodeling involves the incorporation of histone variants that modulate chromatin accessibility for stress-responsive transcription factors. We identify the histone variant H2A.J as a constitutive component of renal TECs, significantly upregulated after exposure to ionizing radiation (IR). Using H2A.J knock-out (KO) mice, we demonstrate that its absence disrupts acute damage responses and prevents coordinated repair, severely impairing regeneration. Mechanistically, H2A.J deficiency compromises mitochondrial function under postirradiation metabolic stress, driving the transition from acute injury to chronic kidney disease via persistent inflammation and maladaptive tubulointerstitial repair. Targeting these epigenetic drivers offers a promising strategy for regenerating damaged kidney tissue in oncology.

molecular biology↗

Micro-CT analyses of the lung in mice: Parameters influencing the radiation dose and acquisition quality

Microcomputed tomography (micro-CT) is a frequently used imaging tool for a wide spectrum of in-vivo mouse models in basic and translational research. To allow an accurate interpretation of micro-CT images, high spatial resolution is necessary. However, this may also lead to a high radiation exposure of the animals. Therefore, animal welfare requires exact information about the expected radiation doses for experimental planning. To gain this, a mouse cadaver was herein used for micro-CT analyses under different conditions. For each radiation dose measurement, the cadaver was labeled with thermoluminescent dosimeter chips around the thoracic skin surface. Micro-CT scans of the thorax were performed with spatial resolutions of 35 {micro}m, 18 {micro}m and 9 {micro}m in combination with Al0.5, Al1.0, CuAl and Cu filters. As a surrogate of image quality, the number of identifiable lung vessels was counted on a transversal micro-CT slice. Measured radiation doses varied from 0.09 Gy up to 5.18 Gy dependent on resolution and filter settings. A significant dose reduction of > 75% was achieved by a Cu filter when compared to an Al0.5 filter. However, this resulted in a markedly reduced image quality and interpretability of microstructures due to higher radiation shielding and lower spatial resolution. Thus, the right combination of distinct filters and several scan protocol settings adjusted to the individual requirements can significantly reduce the radiation dose of micro-CT leading to a higher animal welfare standard.

biophysics↗

Concomitant chemotherapy increases radiotherapy-mediated DNA-damage in peripheral blood lymphocytes

53BP1-foci detection in peripheral blood lymphocytes (PBLs) by immunofluorescence microscopy (IFM) is a sensitive and quantifiable DNA-double-strand-break (DSB) marker. In addition, high-resolution transmission-electron-microscopy (TEM) with immunogold-labeling of 53BP1 and DSB-bound phosphorylated Ku70 (pKu70) can be used to determine the progression of the DNA-repair process. Here, we analyzed whether different modes of irradiation influence the formation of DSBs in the PBLs of patients with cancer, and whether accompanying chemotherapy influences the DSB-appearance. We obtained 86 blood samples before and 0.1, 0.5 and 24 h after irradiation from patients with head and neck, or rectal cancers receiving radiotherapy (RT) or radio-chemotherapy (RCT). 53BP1-foci were quantified by IFM. In addition, TEM was used to quantify gold-labelled pKu70-dimers and 53BP1-clusters within euchromatin and heterochromatin of PBLs. During radiotherapy, persisting 53BP1-foci accumulated in PBLs with increasing numbers of administered RT-fractions. This 53BP1-foci accumulation was not influenced by irradiation technique applied (3D-conformal radiotherapy versus intensity-modulated radiotherapy), dose intensity per fraction, number of irradiation fields, or isodose volume. However, more 53BP1-foci were detected in PBLs of patients treated with accompanying chemotherapy. TEM analyses showed that DSBs, indicated by pKu70, were present for longer periods in PBLs of RCT-patients than in PBLs of RT-only-patients. Moreover, not every residual 53BP1-focus was equivalent to a remaining DSB, since pKu70 was not present at every damage site. Persistent 53BP1-clusters, visualized by TEM, without colocalizing pKu70 likely indicate chromatin alterations after repair completion, or possibly, defective repair. Therefore, IFM-53BP1-foci analyses alone are not adequate to determine individual repair capacity after irradiation of PBLs, as a DSB may be indicated by a 53BP1-focus but not every 53BP1-focus represents a DSB. The level of DNA-damage during RT is influenced by the presence of accompanying chemotherapy.

cell biology↗