Biology subjects

Roorda, M.

Publications and source records attributed to Roorda, M..

6 recordsLinked to original sources

bioRxiv2026

Calorie Restriction-Induced Daily Hibernation in Mice Drives Cyclic DNA Damage and Repair

Hibernation consists of bouts of torpor, characterized by profound decreases in metabolism and body temperature (Tb), alternated with periods of euthermia called interbout arousals, during which normal metabolism and Tb resume. Seasonal hibernators accumulate DNA strand breaks during torpor, which are repaired during arousal. Here, we assess dynamics of DNA damage and repair during serial daily torpor in mice induced by 30% calorie restriction (CR) and investigate the effects of metabolic challenge on DNA repair. Serial daily torpor induced by CR in C57/BL6J mice of both sexes housed at 20{degrees}C lasts 6-12 hours. Like seasonal hibernators, DNA damage increases in CR-induced torpor and is repaired in the subsequent euthermic period, as evidenced by comet assay and {gamma}H2AX accumulation. To metabolically challenge animals, ambient temperature (Ta) was lowered to 4{degrees}C, since torpid mice defend a Tb of around 20{degrees}C or higher. Despite inducing a significant metabolic challenge, housing of torpid mice at 4{degrees}C does not increase DNA damage compared to 20{degrees}C housing. However, reducing Ta to 4{degrees}C during euthermia inhibits DNA repair. Interestingly, p21 levels increase in mice exposed to 4{degrees}C, indicating cell-cycle inhibition during exposure to 4{degrees}C. Thus, 30% CR induces daily cycles of torpor-induced DNA damage and euthermia-associated DNA repair in mice, and exposure to a Ta of 4{degrees}C during arousal inhibits DNA repair mounting a cell cycle inhibition response. Thus, the torpor-arousal cycle may be a contributing factor to the lifespan extension benefits of CR in mice, promoting genomic integrity and thereby cellular and tissue health.

physiology↗

bioRxiv2026

In garden dormouse cerebral cortex, specific transcriptional programs exist for all major phases of hibernation

Hibernators cycle between torpor, a state of profound metabolic and thermoregulatory suppression, and brief arousals during which metabolic rate and body temperature rapidly return to euthermic levels. These repeated physiological pressures require robust mechanisms to preserve brain integrity. Because the cerebral cortex is not thought to control hibernation directly yet must remain viable throughout torpor and recover rapidly during arousal, it provides a useful model for studying neural adaptation to hibernation. We therefore performed RNA sequencing of cerebral cortex from garden dormice (Eliomys quercinus) sampled during summer euthermia (SE), early torpor (TE), late torpor (TL), early arousal (AE), and late arousal (AL). Differential expression analysis revealed strongly stage-specific transcriptional remodeling across the hibernation cycle. Entry into torpor (SE-TE) and the transition from early to late arousal (AE-AL) showed minimal change, with 16 and 2 differentially expressed genes (DEGs), respectively. In contrast, extensive regulation was observed during torpor progression (TE-TL; 576 DEGs) and especially during the transition from late torpor to early arousal (TL-AE; 697 DEGs). Intermediate numbers of DEGs were detected in AL-TE (260) and AL-SE (50). Principal component and enrichment analyses indicated that the dominant axes of variation were associated with RNA processing and proteostatic control, metabolic and redox-related adaptation, and changes in intracellular trafficking and protein handling. In addition, comparison of adjacent contrasts revealed a marked opposite-direction transcriptional reversal between TE-TL and TL-AE, consistent with coordinated reactivation of torpor-associated programs during arousal. Together, these findings support a model in which cortex adaptation to hibernation involves transcriptional reprogramming consistent with metabolic suppression during torpor progression, especially in pathways related to carbohydrate and central carbon metabolism, redox homeostasis, and cellular signalling, followed by rapid reversal of these programs during early arousal.

molecular biology↗

bioRxiv2025

Increased activity of PRMT5-MEP50 complex improves survival of chromosomally unstable cancer cells by increasing tolerance to protein aggregation and proteotoxicity

Most cancers display chromosomal instability (CIN), a condition that leads to increased rates of chromosome missegregation and thus yields aneuploidy. CIN and aneuploidy are detrimental to healthy cells and therefore, aneuploid cells rely on aneuploidy-tolerating mechanisms to adopt a malignant fate. We previously found PRMT5 to be frequently amplified in a mouse model for aneuploid T cell lymphoblastic lymphoma. In this study, we investigated a possible role of PRMT5 as an aneuploidy tolerating gene. We report that PRMT5 is prone to aggregation when expressed at supra-stoichiometric levels compared to its obligate partner protein MEP50 (methylosome protein 50, WDR77). Intriguingly, we also find that protein aggregation, induced by CIN, is mitigated by jointly increased expression of PRMT5 and MEP50. Accordingly, concomitant PRMT5:MEP50 expression renders cancer cells less sensitive to proteasome inhibitors and CIN while inhibition sensitizes cells to CIN. Our findings provide a possible explanation for why PRMT5 and MEP50 display increased expression, particularly in aneuploid cancers and might reveal a targetable vulnerability of aneuploid cancer.

cancer biology↗

bioRxiv2025

WEE1 inhibitors trigger GCN2-mediated activation of the integrated stress response

The WEE1 kinase negatively regulates CDK1/2 to control DNA replication and mitotic entry. Genetic factors that determine sensitivity to WEE1 inhibitors (WEE1i) are largely unknown. A genome-wide insertional mutagenesis screen revealed that mutation of EIF2A, a translation regulator, sensitized to WEE1i. Mechanistically, WEE1i treatment triggers a translational shut-down, which is lethal in combination with the reduced translation of EIF2AKO cells. A genome-wide CRISPR-Cas9 screen revealed that inactivation of integrated stress response (ISR) kinases GCN1/2 rescued WEE1i-mediated cytotoxicity. WEE1i induced GCN2 activation, ATF4 upregulation, and altered ribosome dynamics. Loss of the collided ribosome sensor ZNF598 conversely increased sensitivity to WEE1i. Notably, the ISR was not required for WEE1i to induce DNA damage, premature mitotic entry or sensitization to DNA-damaging chemotherapeutics. ISR activation was independent of CDK1/2 activation. Importantly, WEE1i-mediated ISR activation was independent of WEE1 presence, pointing at off-target effects, which are shared by multiple chemically distinct WEE1i. This response was also observed in peripheral blood mononuclear cells. Importantly, low-dose WEE1 inhibition did not induce ISR activation, while it still synergized with PKMYT1 inhibition. Taken together, WEE1i triggers toxic ISR activation and translational shutdown, which can be prevented by low-dose or combination treatments, while retaining the cell cycle checkpoint-perturbing effects.

cancer biology↗

bioRxiv2021

A synthetic lethal screen identifies HDAC4 as a potential target in MELK overexpressing cancers

Maternal embryonic leucine zipper kinase (MELK) is frequently overexpressed in cancer, but the role of MELK in cancer is still poorly understood. MELK was shown to have roles in many cancer-associated processes including tumor growth, chemotherapy resistance, and tumor recurrence. To determine whether the frequent overexpression of MELK can be exploited in therapy, we performed a high-throughput screen using a library of Saccharomyces cerevisiae mutants to identify genes whose functions become essential when MELK is overexpressed. We identified two such genes: LAG2 and HDA3. LAG2 encodes an inhibitor of the SCF ubiquitin-ligase complex, while HDA3 encodes a subunit of the HDA1 histone deacetylase complex. We find that one of these synthetic lethal interactions is conserved in mammalian cells, as inhibition of a human homolog of HDA3 (HDAC4) is synthetically toxic in MELK overexpression cells. Altogether, our work might provide a new angle of how to exploit MELK overexpression in cancers and might thus lead to novel intervention strategies.

genetics↗

bioRxiv2021

MYC promotes immune-suppression in TNBC via inhibition of IFN signaling

Immune checkpoint inhibitor (ICI) treatment has thus far shown limited efficacy in triple-negative breast cancer (TNBC) patients, presumably due to sparse or unresponsive tumor-infiltrating lymphocytes. We reveal a strong correlation between MYC expression and loss of immune signatures in human TNBC. In mouse models of BRCA1-proficient and -deficient TNBC, MYC overexpression dramatically decreased lymphocyte infiltration in tumors, along with immune signature loss. Likewise, MYC overexpression suppressed inflammatory signaling induced by BRCA1/2 inactivation in human TNBC cell lines. Moreover, MYC overexpression prevented the recruitment and activation of lymphocytes in co-cultures with human and mouse TNBC models. Chromatin immunoprecipitation (ChIP)-sequencing revealed that MYC directly binds promoters of multiple interferon-signaling genes, which were downregulated upon MYC expression. Finally, MYC overexpression suppressed induction of interferon signaling and tumor growth inhibition by a Stimulator of Interferon Genes (STING) agonist. Together, our data reveal that MYC suppresses innate immunity and facilitates immune escape, explaining the poor immunogenicity of MYC-overexpressing TNBCs. Statement of SignificanceMYC suppresses recruitment and activation of immune cells in TNBC by repressing the transcription of interferon genes. These findings provide a mechanistic rationale for the association of high MYC expression levels with immune exclusion in human TNBCs, which might underlie the relatively poor response of many TNBCs to ICI.

cancer biology↗