Biology subjects

Paquette, M.

Publications and source records attributed to Paquette, M..

11 recordsLinked to original sources

bioRxiv2021

AMPK-dependent phosphorylation is required for transcriptional activation of TFEB/TFE3

Increased autophagy and lysosomal activity promote tumor growth, survival and chemo-resistance. During acute starvation, autophagy is rapidly engaged by AMPK activation and mTORC1 inhibition to maintain energy homeostasis and cell survival. TFEB and TFE3 are master transcriptional regulators of autophagy and lysosomal activity and their cytoplasm/nuclear shuttling is controlled by mTORC1-dependent multisite phosphorylation. However, it is not known whether and how the transcriptional activity of TFEB or TFE3 is regulated. We show that AMPK mediates phosphorylation of TFEB and TFE3 on three serine residues, leading to TFEB/TFE3 transcriptional activity upon nutrient starvation, FLCN depletion and pharmacological manipulation of mTORC1 or AMPK. AMPK loss does not affect TFEB/TFE3 nuclear localization nor protein levels but reduces their transcriptional activity. Collectively, we show that mTORC1 specifically controls TFEB/TFE3 cytosolic retention whereas AMPK is essential for TFEB/TFE3 transcriptional activity. This dual and opposing regulation of TFEB/TFE3 by mTORC1 and AMPK is reminiscent of the regulation of another critical regulator of autophagy, ULK1. Surprisingly, we show that chemoresistance is mediated by AMPK-dependent activation of TFEB, which is abolished by pharmacological inhibition of AMPK or mutation of serine 466/467/469 to alanine residues within TFEB. Altogether, we show that AMPK is a key regulator of TFEB/TFE3 transcriptional activity, and we validate AMPK as a promising target in cancer therapy to evade chemotherapeutic resistance.

cancer biology↗

bioRxiv2020

Axon Diameter Measurements using Diffusion MRI are Infeasible

The feasibility of non-invasive axonal diameter quantification with diffusion MRI is a strongly debated topic due to the neuroscientific potential of such information and its relevance for the axonal signal transmission speed. It has been shown that under ideal conditions, the minimal diameter producing detectable signal decay is bigger than most human axons in the brain, even using the strongest currently available MRI systems. We show that resolving the simplest situations including multiple diameters is unfeasible even with diameters much bigger than the diameter limit. Additionally, the recently proposed effective diameter resulting from fitting a single value over a distribution is almost exclusively influenced by the biggest axons. We show how impractical this metric is for comparing different distributions. Overall, axon diameters currently cannot be quantified by diffusion MRI in any relevant way.

biophysics↗

bioRxiv2020

FLCN Gene Ablation Reduces Fibrosis and Inflammation in a Diet-Induced NASH Model

Non-alcoholic steatohepatitis (NASH) represents a major economic burden and is characterized by triglyceride accumulation, inflammation, and fibrosis. No pharmacological agents are currently approved to treat this condition. Emerging data suggests an important role of autophagy in this condition, which serves to degrade intracellular lipid stores, reduce hepatocellular damage, and dampen inflammation. Autophagy is primarily regulated by the transcription factors TFEB and TFE3, which are negatively regulated by mTORC1. Given that FLCN is an mTORC1 activator via its GAP activity towards RagC/D, we generated a liver specific Flcn knockout mouse model to study its role in NASH progression. We demonstrate that loss of FLCN results in reduced triglyceride accumulation, fibrosis, and inflammation in mice exposed to a NASH-inducing diet. Hence, the GAP activity of FLCN could a promising target for small molecule drugs to treat NASH progression by specifically activating autophagy and lysosomal biogenesis while leaving mRNA translation machinery unperturbed. Collectively, these results show an unexpected role for FLCN in NASH progression and highlight new possibilities for treatment strategies through its role in hepatocyte homeostasis.

pathology↗

bioRxiv2020

Increased Sensitivity and Signal-to-Noise Ratio in Diffusion-Weighted MRI using Multi-Echo Acquisitions

Post-mortem diffusion MRI (dMRI) enables acquisitions of structural imaging data with otherwise unreachable resolutions - at the expense of longer scanning times. These data are typically acquired using highly segmented image acquisition strategies, thereby resulting in an incomplete signal decay before the MRI encoding continues. Especially in dMRI, with low signal intensities and lengthy contrast encoding, such temporal inefficiency translates into reduced image quality and longer scanning times. This study introduces Multi Echo (ME) acquisitions to dMRI on a human MRI system - a time-efficient approach, which increases SNR (Signal-to-Noise Ratio) and reduces noise bias for dMRI images. The benefit of the introduced ME dMRI method was validated using numerical Monte Carlo simulations and showcased on a post-mortem brain of a wild chimpanzee. The proposed Maximum Likelihood Estimation echo combination results in an optimal SNR without detectable signal bias. The combined strategy comes at a small price in scanning time (here 30% additional) and leads to a substantial SNR increase (here up to 1.9x which is equivalent to 3.6 averages) and a general reduction of the noise bias.

neuroscience↗

bioRxiv2019

Dynamic cerebral autoregulation of endurance-trained men following 6 weeks of high-intensity interval training to exhaustion

Elevated cardiorespiratory fitness (CRF) is associated with reduced dynamic cerebral autoregulation (dCA), but the impact of exercise training per se on dCA remains equivocal. In addition, resting cerebral blood flow (CBF) and dCA after high-intensity interval training (HIIT) in individuals with already high CRF is unknown. We examined to what extent 6 weeks of HIIT affect resting CBF and dCA in cardiorespiratory fit men and explored if potential changes are intensity-dependent. Endurance-trained men were assigned to group HIIT85 (85% of maximal aerobic power, 1 to 7 min effort bouts, n = 8) and HIIT115 (115% of maximal aerobic power, 30 s to 1 min effort bouts, n = 9). Training sessions were completed until exhaustion 3 times/week over 6 weeks. Mean arterial pressure (MAP) and middle cerebral artery mean blood velocity (MCAvmean) were measured continuously at rest and during repeated squat-stands (0.05 and 0.10 Hz). Transfer function analysis (TFA) was used to characterize dCA on driven blood pressure oscillations during repeated squat-stands. Neither training nor intensity had an effect on resting MAP and MCAvmean (both P > 0.05). TFA phase during 0.10 Hz squat-stands decreased after HIIT irrespective of intensity (HIIT85: 0.77 {+/-} 0.22 vs. 0.67 {+/-} 0.18 radians; HIIT115: pre: 0.62 {+/-} 0.19 vs. post: 0.59 {+/-} 0.13 radians, time effect P = 0.048). These results suggest that HIIT over 6 weeks have no apparent benefits on resting CBF, but a subtle attenuation in dCA is seen posttraining irrespective of intensity training in endurance-trained men.\n\nNEW & NOTEWORTHYThe novel findings of this study are that 6 weeks of submaximal and supramaximal high-intensity interval exercise to exhaustion reduce dynamic cerebral autoregulation irrespective of training intensity in endurance-trained men. However, these HIIT protocols do not influence resting cerebral blood flow in these individuals. The results indicate the cerebrovasculature of endurance-trained men has an attenuated ability to react to large and rapid changes in blood pressure following HIIT.

physiology↗

bioRxiv2019

Cardiac remodeling after six weeks of high-intensity interval training to exhaustion in endurance-trained males

High-intensity interval training (HIIT) improves physical performance of endurance athletes, although studies examining its cardiovascular effects are sparse. We evaluated the impact of HIIT on blood pressure, heart rate and cardiac cavities size and function in endurance-trained adults. Seventeen endurance-trained males underwent 24-hour ambulatory blood pressure monitoring (ABPM) and Doppler echocardiography at baseline and after 6 weeks of HIIT. Participants were divided in 2 groups (85% maximal aerobic power (MP); HIIT85, n=8 and 115% MP; HIIT115, n = 9) in order to compare the impact of different HIIT intensities. ABPM and cardiac chambers size and function were similar between groups at baseline. HIIT reduced heart rate (55 {+/-} 8 vs. 51 {+/-} 7 bpm; p= 0.003), systolic blood pressure (SBP; 121 {+/-} 11 vs 118 {+/-} 9 mmHg; p = 0.01), mean arterial pressure (MAP; 90 {+/-} 8 vs 89 {+/-} 6 mmHg; p = 0.03) and pulse pressure (52 {+/-} 6 vs 49 {+/-} 5 mmHg; p = 0.01), irrespective of training intensity. Left atrium (LA) volumes increased after HIIT (maximal: 50 {+/-} 14 vs 54 {+/-}14 ml; p= 0.02; minimal: 15 {+/-} 5 vs 20 {+/-} 8 ml; p = 0.01) in both groups. Right ventricle (RV) global longitudinal strain lowered after training in the HIIT85 group only (20 {+/-} 4 vs. 17 {+/-} 3%, p = 0.04). In endurance-trained males, six weeks of HIIT reduce SBP and MAP and increase LA volumes irrespective of training intensity, whereas submaximal HIIT deteriorates RV systolic function.\n\nKey pointsO_LIHigh-intensity interval training improves physical performance of endurance athletes, although studies examining its cardiovascular effects are sparse.\nC_LIO_LIWe evaluated the impact of submaximal (85% maximal aerobic power) and supramaximal (115% maximal aerobic power) high-intensity interval training on ambulatory blood pressure, heart rate and cardiac cavities size and function in endurance-trained adults.\nC_LIO_LIIrrespective of training intensity, six weeks of high-intensity interval training increase left atrial volumes in endurance-trained adults, whereas the submaximal training decreases right ventricular systolic function.\nC_LIO_LIThese results may help identify the exercise threshold for potential toxicity of intense exercise training for at-risk individuals and ideal exercise training regimens conferring optimal cardiovascular protection and adapted endurance training for athletes.\nC_LI

physiology↗

bioRxiv2018

Tractography Reproducibility Challenge with Empirical Data (TraCED): The 2017 ISMRM Diffusion Study Group Challenge

PurposeFiber tracking with diffusion weighted magnetic resonance imaging has become an essential tool for estimating in vivo brain white matter architecture. Fiber tracking results are sensitive to the choice of processing method and tracking criteria. Phantom studies provide concrete quantitative comparisons of methods relative to absolute ground truths, yet do not capture variabilities because of in vivo physiological factors.\n\nMethodsTo date, a large-scale reproducibility analysis has not been performed for the assessment of the newest generation of tractography algorithms with in vivo data. Reproducibility does not assess the validity of a brain connection however it is still of critical importance because it describes the variability for an algorithm in group studies. The ISMRM 2017 TraCED challenge was created to fulfill the gap. The TraCED dataset consists of a single healthy volunteer scanned on two different scanners of the same manufacturer. The multi-shell acquisition included b-values of 1000, 2000 and 3000 s/mm2 with 20, 45 and 64 diffusion gradient directions per shell, respectively.\n\nResultsNine international groups submitted 46 tractography algorithm entries. The top five submissions had high ICC > 0.88. Reproducibility is high within these top 5 submissions when assessed across sessions or across scanners. However, it can be directly attributed to containment of smaller volume tracts in larger volume tracts. This holds true for the top five submissions where they are contained in a specific order. While most algorithms are contained in an ordering there are some outliers.\n\nConclusionThe different methods clearly result in fundamentally different tract structures at the more conservative specificity choices (i.e., volumetrically smaller tractograms). The data and challenge infrastructure remain available for continued analysis and provide a platform for comparison.

neuroscience↗

bioRxiv2018

The transcription factors TFEB and TFE3 link the FLCN-AMPK signaling axis to innate immune response and pathogen resistance

TFEB and TFE3 are transcriptional regulators of the innate immune response, but the mechanisms regulating their activation upon pathogen infection are poorly elucidated. Using C. elegans and mammalian models, we report that the master metabolic modulator 5-AMP-activated protein kinase (AMPK) and its negative regulator Folliculin (FLCN) act upstream of TFEB/TFE3 in the innate immune response, independently of the mTORC1 signaling pathway. In nematodes, loss of FLCN or overexpression of AMPK conferred pathogen resistance via activation of TFEB/TFE3-dependent antimicrobial genes, while ablation of total AMPK activity abolished this phenotype. Similarly, in mammalian cells, loss of FLCN or pharmacological activation of AMPK induced TFEB/TFE3-dependent pro-inflammatory cytokine expression. Importantly, a rapid reduction in cellular ATP levels in murine macrophages was observed upon lipopolysaccharide (LPS) treatment accompanied by an acute AMPK activation and TFEB nuclear localization. These results uncover an ancient, highly conserved and pharmacologically actionable mechanism coupling energy status with innate immunity.

immunology↗

bioRxiv2018

Functional implications of impaired dynamic cerebral autoregulation in young healthy women; a comparative investigation

Women exhibit higher prevalence of orthostatic hypotension with presyncopal symptoms compared to men. These symptoms could be influenced by an attenuated ability of the cerebrovasculature to respond to rapid changes in blood pressure (BP) [dynamic cerebral autoregulation (dCA)]. However, the influence of sex on dCA remains equivocal. We compared dCA in 11 women (25 {+/-} 2 y) and 11 age-matched men (24 {+/-} 1 y) using a multimodal approach including a sit-to-stand maneuver and forced oscillations (5 min of squat-stand performed at 0.05 and 0.10 Hz). The prevalence of initial orthostatic hypotension (IOH; decrease in systolic [≥] 40 mmHg and/or diastolic BP [≥] 20 mmHg) during the first 15 sec of sit-to-stand was determined as a functional outcome. In women, the decrease in mean middle cerebral artery blood velocity (MCAvmean) following the sit-to-stand was greater (-20 {+/-} 8 vs. -11 {+/-} 7 cm sec-1; p=0.018) and the onset of the regulatory change (time lapse between the beginning of the sit-to-stand and the increase in the conductance index (MCAvmean/mean arterial pressure(MAP)) was delayed (p=0.007). Transfer function analysis gain during 0.05 Hz squat-stand was [~]48% higher in women (6.4 {+/-} 1.3 vs. 3.8 {+/-} 2.3 sec; p=0.017). The prevalence of IOH was comparable between groups (4/9 vs. 5/9, p=0.637). These results indicate the cerebrovasculature of healthy women has an attenuated ability to react to large and rapid changes in BP in the face of preserved orthostasis, which could be related to a higher cerebrovascular reserve to face a rapid transient hypotension.\n\nNEWS & NOTEWORTHYThe novel findings of this study are that healthy women have impaired dynamic cerebral autoregulation, although the prevalence of orthostatic intolerance was similar in women and men. These results indicate the cerebrovasculature of healthy women has an attenuated ability to react to large and rapid changes in blood pressure in the face of preserved orthostasis, which could be related to a higher cerebrovascular reserve to face a rapid transient hypotension.

physiology↗

bioRxiv2017

Diminished dynamic cerebral autoregulatory capacity with forced oscillations in mean arterial pressure with elevated cardiorespiratory fitness

The effect that cardiorespiratory fitness has on the dynamic cerebral autoregulatory capacity during changes in mean arterial pressure (MAP) remains equivocal. Using a multiple-metrics approach, challenging MAP across the spectrum of physiological extremes (i.e. spontaneous through forced MAP oscillations), we characterized dynamic cerebral autoregulatory capacity in 19 male endurance athletes and 8 controls via three methods: [1] onset of regulation (i.e., time delay before an increase in middle cerebral artery (MCA) conductance [MCA flow velocity (MCAv)/mean arterial pressure (MAP)] and rate of regulation, after transient hypotension induced by sit-to-stand, and transfer function analysis (TFA) of MAP and MCAv responses during [2] spontaneous and [3] forced oscillations (5-min of squat-stand maneuvers performed at 0.05 and 0.10 Hz). Reductions in MAP and mean MCAv (MCAVmean) during initial orthostatic stress (0-30 s after sit-to-stand) and the prevalence of orthostatic hypotension were also determined. Onset of regulation was delayed after sit-to-stand in athletes (3.1{+/-}1.7 vs. 1.5{+/-}1.0 s; p=0.03), but rate of regulation was not different between groups (0.24{+/-}0.05 vs. 0.21{+/-}0.09 s-1; p=0.82). While both groups had comparable TFA metrics during spontaneous oscillations, athletes had higher TFA gain during 0.10 Hz squat-stand vs. recreational controls (p=0.01). Reductions in MAP (p=0.15) and MCAVmean (p=0.11) during orthostatic stress and the prevalence of initial orthostatic hypotension (p=0.65) were comparable between groups. These results indicate an intact ability of the cerebral vasculature to react to spontaneous oscillations but an attenuated capability to counter rapid and large changes in MAP in individuals with elevated cardiorespiratory fitness.\n\nNew findingsO_ST_ABSWhat is the central question of this study?C_ST_ABSO_LIWhat is the impact of cardiorespiratory fitness on dynamic cerebral autoregulatory capacity?\nC_LI\n\nWhat is the main finding and its importance?O_LIOur results indicate an intact ability of the cerebral vasculature to react to spontaneous oscillations but an attenuated capability to counter rapid and large changes in mean arterial pressure in individuals with elevated cardiorespiratory fitness.\nC_LI

physiology↗

bioRxiv2016

Tractography-based connectomes are dominated by false-positive connections

Fiber tractography based on non-invasive diffusion imaging is at the heart of connectivity studies of the human brain. To date, the approach has not been systematically validated in ground truth studies. Based on a simulated human brain dataset with ground truth white matter tracts, we organized an open international tractography challenge, which resulted in 96 distinct submissions from 20 research groups. While most state-of-the-art algorithms reconstructed 90% of ground truth bundles to at least some extent, on average they produced four times more invalid than valid bundles. About half of the invalid bundles occurred systematically in the majority of submissions. Our results demonstrate fundamental ambiguities inherent to tract reconstruction methods based on diffusion orientation information, with critical consequences for the approach of diffusion tractography in particular and human connectivity studies in general.

neuroscience↗