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Chen, J. J.

Publications and source records attributed to Chen, J. J..

52 records · Page 3Linked to original sources

Generating dynamic carbon-dioxide from the respiratory-volume time series: A feasibility study using neural networks

In the context of fMRI, carbon dioxide (CO2) is a well-known vasodilator that has been widely used to monitor and interrogate vascular physiology. Moreover, spontaneous fluctuations in end-tidal carbon dioxide (PETCO2) reflects changes in arterial CO2 and has been demonstrated as the largest physiological noise source in the low-frequency range of the resting-state fMRI (rs-fMRI) signal. Increasing appreciation for the role of CO2 in fMRI has given rise to methods that use it for physiological denoising or estimating cerebrovascular reactivity. However, the majority of rs-fMRI studies do not involve CO2 recordings, and most often only heart rate and respiration are recorded. While the intrinsic link between these latter metrics and CO2 led to suggested possible analytical models, they have not been widely applied. In this proof-of-concept study, we propose a deep learning approach to reconstruct CO2 and PETCO2 data from respiration waveforms in the resting state. We demonstrate that the one-to-one mapping between respiration and CO2 recordings can be well predicted using fully convolutional networks (FCNs), achieving a Pearson correlation coefficient (r) of 0.946 {+/-} 0.056 with the ground truth CO2. Moreover, dynamic PETCO2 can be successfully derived from the predicted CO2, achieving r of 0.512 {+/-} 0.269 with the ground truth. Importantly, the FCN-based methods outperform previously proposed analytical methods. In addition, we provide guidelines for quality assurance of respiration recordings for the purposes of CO2 prediction. Our results demonstrate that dynamic CO2 can be obtained from respiration-volume using neural networks, complementing the still few reports in deep-learning of physiological fMRI signals, and paving the way for further research in deep-learning based bio-signal processing.

bioinformatics↗

Modeling the carbon-dioxide response function in fMRI under task and resting-state conditions

Conventionally, cerebrovascular reactivity (CVR) is estimated as the amplitude of the hemodynamic response to vascular stimuli. While the CVR amplitude has established clinical utility, the temporal characteristics of CVR have been increasingly explored and may yield even more pathology-sensitive parameters. This work is motivated by the current need to evaluate the feasibility of dCVR modeling in various noise conditions. In this work, we present a comparison of several recently published model-based deconvolution approaches for estimating h(t), including maximum a posterior likelihood (MAP), inverse logit (IL), canonical correlation analysis (CCA), and basis expansion (using Gamma and Laguerre basis sets). To aid the comparison, we devised a novel simulation framework that allowed us to target a wide range of SNRs, ranging from 10 to -7 dB, representative of both task and resting-state CO2 changes. In addition, we built ground-truth h(t) into our simulation framework, overcoming the practical limitation that the true h(t) is unknown in methodological evaluations. Moreover, to best represent realistic noise found in fMRI scans, we extracted it from in-vivo resting-state scans. Furthermore, we introduce a simple optimization of the CCA method (CCAopt) and compare its performance to these existing methods. Our findings suggest that model-based methods can reasonably estimate dCVR even amidst high noise, and in a manner that is largely independent of the underlying model assumptions for each method. We also provide a quantitative basis for making methodological choices, based on the desired dCVR parameters, the estimation accuracy and computation time. The BEL method provided the highest accuracy and robustness, followed by the CCAopt and IL methods. Of the three, the CCAopt method required the lowest computational time. These findings lay the foundation for wider adoption of dCVR estimation in CVR mapping.

biophysics↗

White matter microstructural integrity across the adult lifespan: Combined perspective of diffusion tensor and kurtosis imaging

Studies of healthy brain aging have reported diffusivity patterns associated with white matter degeneration using diffusion tensor imaging (DTI), which assumes that diffusion measured at the typical b-value (approximately 1000 s/mm2) is Gaussian. Diffusion kurtosis imaging (DKI) is an extension of DTI that measures non-Gaussian diffusion (kurtosis) to better capture microenvironmental changes by incorporating additional data at a higher b-value. In this study, using UK Biobank data (b values of 1000 and 2000 s/mm2), we investigate (1) the extent of novel information gained from adding diffusional kurtosis to diffusivity observations in aging, and (2) how conventional DTI metrics in aging compare with diffusivity metrics derived from DKI, which are corrected for kurtosis. We find a general pattern of lower kurtosis alongside higher diffusivity among older adults. We also find differences between diffusivity metrics derived from DTI and DKI, emphasizing the importance of accounting for non-Gaussian diffusion. This work highlights the utility of measuring diffusional kurtosis as a simple addition to conventional diffusion imaging of aging.

neuroscience↗

Performance of temporal and spatial ICA in identifying and removing low-frequency physiological and motion effects in resting-state fMRI

Effective separation of signal from noise (including physiological processes and head motion) is one of the chief challenges for improving the sensitivity and specificity of resting-state fMRI (rs-fMRI) measurements and has a profound impact when these noise sources vary between populations. Independent component analysis (ICA) is an approach for addressing these challenges. Conventionally, due to the lower amount of temporal than spatial information in rs-fMRI data, spatial ICA (sICA) is the method of choice. However, with recent developments in accelerated fMRI acquisitions, the temporal information is becoming enriched to the point that the temporal ICA (tICA) has become more feasible. This is particularly relevant as physiological processes and motion exhibit very different spatial and temporal characteristics when it comes to rs-fMRI applications, leading us to conduct a comparison of the performance of sICA and tICA in addressing these types of noise. In this study, we embrace the novel practice of using theory (simulations) to guide our interpretation of empirical data. We find empirically that sICA can identify more noise-related signal components than tICA. However, on the merit of functional-connectivity results, we find that while sICA is more adept at reducing whole-brain motion effects, tICA performs better in dealing with physiological effects. These interpretations are corroborated by our simulation results. The overall message of this study is that if ICA denoising is to be used for rs-fMRI, there is merit in considering a hybrid approach in which physiological and motion-related noise are each corrected for using their respective best-suited ICA approach. Impact StatementResting-state fMRI is influenced by low-frequency physiological noise and head motion. Independent component analysis (ICA) is becoming increasingly relied on for reducing these influences, but the utility of spatial and temporal ICA remains unclear. We conducted a comparison of the performance of these two ICA types, using physiological-noise and motion time courses as reference. We found that spatial ICA is more adept at reducing motion effects, while temporal ICA performs better in dealing with physiological effects. We believe these findings provide much-needed clarity on the role of ICA, and recommend using a hybrid of tICA and sICA as a paradigm shift in resting-state fMRI.

neuroscience↗

Exo1-protected DNA nicks direct crossover formation in meiosis

In most sexually reproducing organisms crossing over between chromosome homologs during meiosis is critical for the viability of haploid gametes. Most crossovers that form in meiosis in budding yeast result from the biased resolution of double Holliday Junction (dHJ) intermediates. This dHJ resolution step involves the actions Rad2/XPG family nuclease Exo1 and the Mlh1-Mlh3 mismatch repair endonuclease. At present little is known about how these factors act in meiosis at the molecular level. Here we show that Exo1 promotes meiotic crossing over by protecting DNA nicks from ligation. We found that structural elements in Exo1 required for interactions with DNA, such as bending of DNA during nick/flap recognition, are critical for its role in crossing over. Consistent with these observations, meiotic expression of the Rad2/XPG family member Rad27 partially rescued the crossover defect in exo1 null mutants, and meiotic overexpression of Cdc9 ligase specifically reduced the crossover levels of exo1 DNA binding mutants to levels approaching the exo1 null. In addition, our work identified a role for Exo1 in crossover interference that appears independent of its resection activity. Together, these studies provide experimental evidence for Exo1-protected nicks being critical for the formation of meiotic crossovers and their distribution.

molecular biology↗

Dopamine D1 receptor activation and cAMP/PKA signalling mediate Brd4 recruitment to chromatin to regulate gene expression in rat striatal neurons

The activity of striatal medium-spiny projection neurons is regulated by dopamine acting principally at D1 and D2 dopamine receptors. The dopamine D1 receptor (D1R) is a Gs/olf-coupled GPCR which activates a cAMP/PKA/DARPP-32 signalling cascade that increases excitability and facilitates plasticity, partly through the regulation of transcription. Transcriptional regulation downstream of the D1R involves the activation of PKA, which can translocate to the nucleus to phosphorylate various targets. The chromatin reader Brd4 regulates transcription induced by neurotrophic factors in cortical neurons and has also been implicated in dopamine-dependent striatal functions. Brd4 is activated by phosphorylation; this facilitates its binding to acetylated histones at promoters and enhancers. In non-neuronal cells, Brd4 is recruited to chromatin in response to PKA signalling. However, it is unknown whether Brd4 is involved in transcriptional activation by the D1R in striatal neurons. Here, we demonstrate that cAMP/PKA signalling increases Brd4 recruitment to dopamine-induced genes in striatal neurons, and that knockdown or inhibition of Brd4 modulated D1R-induced gene expression. Specifically, inhibition of Brd4 with the bromodomain inhibitor JQ1 suppressed the expression of [~]25% of D1R-upregulated genes, while increasing the expression of a subset of immediate-early genes, including Fos and Jun. This pro-transcriptional effect of JQ1 was P-TEFb-dependent, and mediated through inhibition of the BD1 bromodomain of Brd4. Finally, we report that JQ1 treatment downregulated expression of many GPCRs and also impaired ERK1/2 signalling in striatal neurons. Our findings identify Brd4 as a novel regulator of D1R-dependent transcription and delineate complex bi-directional effects of bromodomain inhibitors on neuronal transcription.

neuroscience↗

Vascular origins of low-frequency oscillations in the cerebrospinal fluid signal in resting-state fMRI: Interpretation using photoplethysmography

Slow and rhythmic spontaneous oscillations of cerebral blood flow are well known to have diagnostic utility, notably frequencies of 0.008-0.03 Hz (B-waves) and 0.05-0.15Hz (Mayer waves or M waves). However, intracranial measurements of these oscillations have been difficult. Oscillations in the cerebrospinal fluid (CSF), which are influenced by the cardiac pulse wave, represent a possible avenue for non-invasively tracking these oscillations using resting-state functional MRI (rs-fMRI), and have been used to correct for vascular oscillations in rs-fMRI functional connectivity calculations. However, the relationship between low-frequency CSF and vascular oscillations is unclear. In this study, we investigate this relationship using fast simultaneous multi-slice rs-fMRI coupled with fingertip photoplethysmography (PPG). We not only extract B-wave and M-wave range spectral power from the PPG signal, but also derive the pulse-intensity ratio (PIR, a surrogate of slow blood-pressure oscillations), the second-derivative of the PPG (SDPPG, a surrogate of arterial stiffness) and heart-rate variability (HRV). The main findings of this study are: (1) signals in different CSF regions (ROIs) are not equivalent in their vascular contributions or in their associations with vascular and tissue rs-fMRI signals; (2) the PPG signal contains the highest signal contribution from the M-wave range, while PIR contains the highest signal contribution from the B-wave range; (3) in the low-frequency range, PIR is more strongly associated with rs-fMRI signal in the CSF than PPG itself, and than HRV and SDPPG; (4) PPG-related vascular oscillations only contribute to < 20% of the CSF signal in rs-fMRI, insufficient support for the assumption that low-frequency CSF signal fluctuations directly reflect vascular oscillations. These findings caution the use of CSF as a monolithic region for extracting physiological nuisance regressors in rs-fMRI applications. They also pave the way for using rs-fMRI in the CSF as a potential tool for tracking cerebrovascular health through, for instance the strong relationship between PIR and the CSF signal.

physiology↗

Liver x receptor alpha drives chemoresistance in response to side-chain hydroxycholesterols in triple negative breast cancer

Triple negative breast cancer (TNBC) is challenging to treat successfully because targeted therapies do not exist. Instead, systemic therapy is typically restricted to cytotoxic chemotherapy, which fails more often in patients with elevated circulating cholesterol. Liver x receptors are ligand-dependent transcription factors that are homeostatic regulators of cholesterol, and are linked to regulation of broad-affinity xenobiotic transporter activity in non-tumor tissues. We show that LXR ligands confer chemotherapy resistance in TNBC cell lines and xenografts, and that LXRalpha is necessary and sufficient to mediate this resistance. Furthermore, in TNBC patients who had cancer recurrences, LXRalpha and ligands were independent markers of poor prognosis and correlated with P-glycoprotein expression. However, in patients who survived their disease, LXRalpha signaling and P-glycoprotein were decoupled. These data reveal a novel chemotherapy resistance mechanism in this poor prognosis subtype of breast cancer. We conclude that systemic chemotherapy failure in some TNBC patients is caused by co-opting the LXRalpha:P-glycoprotein axis, a pathway highly targetable by therapies that are already used for prevention and treatment of other diseases.

cancer biology↗

Spt5 phosphorylation and the Rtf1 Plus3 domain promote Rtf1 function through distinct mechanisms

Rtf1 is a conserved RNA polymerase II (RNAPII) elongation factor that promotes co-transcriptional histone modification, RNAPII transcript elongation, and mRNA processing. Rtf1 function requires phosphorylation of Spt5, an essential RNAPII processivity factor. Spt5 is phosphorylated within its C-terminal domain (CTD) by cyclin-dependent kinase 9 (Cdk9), catalytic component of positive transcription elongation factor b (P-TEFb). Rtf1 recognizes phosphorylated Spt5 (pSpt5) through its Plus3 domain. Since Spt5 is a unique target of Cdk9, and Rtf1 is the only known pSpt5-binding factor, the Plus3/pSpt5 interaction is thought to be a key Cdk9-dependent event regulating RNAPII elongation. Here we dissect Rtf1 regulation by pSpt5 in the fission yeast Schizosaccharomyces pombe. We demonstrate that the Plus3 domain of Rtf1 (Prf1 in S. pombe) and pSpt5 are functionally distinct, and that they act in parallel to promote Prf1 function. This alternate Plus3 domain function involves an interface that overlaps with the pSpt5 binding site and that can interact with single-stranded nucleic acid or with the Polymerase Associated Factor (PAF) Complex in vitro. We further show that the C-terminal region of Prf1, which also interacts with PAF, has a similar parallel function with pSpt5. Our results elucidate unexpected complexity underlying Cdk9-dependent pathways that regulate transcription elongation.

molecular biology↗

Controlling for the effect of arterial-CO2 fluctuations in resting-state fMRI: Comparing end-tidal CO2 clamping and retroactive CO2 correction

The BOLD signal, as the basis of functional MRI, arises from both neuronal and vascular factors, with their respective contributions to resting state-fMRI still unknown. Among the factors contributing to "physiological noise", dynamic arterial CO2 fluctuations constitutes the strongest and the most widespread modulator of the grey-matter rs-fMRI signal. Some important questions are: (1) if we were able to clamp arterial CO2 such that fluctuations are removed, what would happen to rs-fMRI measures? (2) falling short of that, is it possible to retroactively correct for CO2 effects with equivalent outcome? In this study 13 healthy subjects underwent two rs-fMRI acquisition: During the "clamped" run, end-tidal CO2 (PETCO2) is clamped to the average PETCO2 level of each participant, while during the "free-breathing" run, the PETCO2 level is passively monitored but not controlled. PETCO2 correction was applied to the free-breathing data by convolving PETCO2 with its BOLD response function, and then regressing out the result. We computed the BOLD resting-state fluctuation amplitude (RSFA), as well as seed-independent mean functional connectivity (FC) as the weighted global brain connectivity (wGBC). Furthermore, connectivity between conditions were compared using coupled intrinsic-connectivity distribution (ICD) method. We ensured that PETCO2 clamping did not significantly alter heart-beat and respiratory variation. We found that neither PETCO2 clamping nor correction produced significant change in RSFA and wGBC. In terms of the ICD, PETCO2 clamping and correction both reduced FC strength in the majority of grey matter regions, although the effect of PETCO2 correction is considerably smaller than the effect of PETCO2 clamping. Furthermore, while PETCO2 clamping reduced inter-subject variability in FC, PETCO2 correction increased the variability. Overall PETCO2 correction is not the equivalent of PETCO2 clamping, although it shifts FC values towards the same direction as clamping does.

neuroscience↗

Suppressors of YpsA-mediated cell division inhibition in Bacillus subtilis

Although many bacterial cell division factors have been uncovered over the years, evidence from recent studies points to the existence of yet to be discovered factors involved in cell division regulation. Thus, it is important to identify factors and conditions that regulate cell division to obtain a better understanding of this fundamental biological process. We recently reported that in the Gram-positive organisms Bacillus subtilis and Staphylococcus aureus, increased production of YpsA resulted in cell division inhibition. In this study, we isolated spontaneous suppressor mutations to uncover critical residues of YpsA and the pathways through which YpsA may exert its function. Using this technique, we were able to isolate four unique intragenic suppressor mutations in ypsA (E55D, P79L, R111P, G132E) that rendered the mutated YpsA non-toxic upon overproduction. We also isolated an extragenic suppressor mutation in yfhS, a gene that encodes a protein of unknown function. Subsequent analysis confirmed that cells lacking yfhS were unable to undergo filamentation in response to YpsA overproduction. We also serendipitously discovered that YfhS may play a role in cell size regulation. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=193 SRC="FIGDIR/small/946632v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@1098d63org.highwire.dtl.DTLVardef@216f86org.highwire.dtl.DTLVardef@121050eorg.highwire.dtl.DTLVardef@d2ef3a_HPS_FORMAT_FIGEXP M_FIG C_FIG ABBREVIATED SUMMARYIn Bacillus subtilis, we discovered that increased expression of ypsA results in cell division inhibition and impairment of colony formation on solid medium. Colonies that do arise possess compensatory suppressor mutations. Analysis of one such suppressor mutation led us to a protein of unknown function, YfhS, which appears to play a role in regulating cell length and cell width.

microbiology↗

Compromised function of an ESCRT complex promotes endolysosomal escape of tau seeds and propagation of tau aggregation

Intercellular propagation of protein aggregation is emerging as a key mechanism in the progression of several neurodegenerative diseases, including Alzheimers Disease and frontotemporal dementia. However, we lack a systematic understanding of the cellular path-ways controlling prion-like propagation. To uncover such pathways, we performed CRISPR interference (CRISPRi) screens in a human cell-based model of propagation of tau aggregation. Our screens uncovered that knockdown of several components of the ESCRT machinery, including CHMP6, or CHMP2A in combination with CHMP2B (a gene linked to familial fronto-temporal dementia), promote propagation of tau aggregation. We found that knockdown of these genes caused damage to endolysosomal membranes, consistent with a role for the ESCRT pathway in endolysosomal membrane repair. Leakiness of the endolysosomal compartment significantly enhanced prion-like propagation of tau aggregation, likely by making tau seeds more available to pools of cytoplasmic tau. Together, these findings suggest that endolysosomal escape is a critical step in tau propagation.

cell biology↗

Magnetic Resonance Fingerprinting with Combined Gradient- and Spin-echo Echo-planar Imaging: Simultaneous Estimation of T1, T2 and T2* with integrated-B1 Correction

PurposeTo introduce a novel magnetic-resonance fingerprinting (MRF) framework with single-shot echo-planar imaging (EPI) readout to simultaneously estimate tissue T2, T1 and T2*, and integrate B1 correction.\n\nMethodsSpin-echo EPI is combined with gradient-echo EPI to achieve T2 estimation as well as T1 and T2* quantification. In the dictionary matching step, the GE-EPI data segment provides estimates of tissue T1 and T2* with additional B1 information, which are then incorporated into the T2-matching step that uses the SE-EPI data segment. In this way, biases in T2 and T2* estimates do not affect each other.\n\nResultsAn excellent correspondence was found between our T1, T2, and T2* estimates and results obtained from standard approaches in both phantom and human scans. In the phantom scan, a linear relationship with R2>0.96 was found for all parameter estimates. The maximum error in the T2 estimate was found to be below 6%. In the in-vivo scan, similar contrast was noted between MRF and standard approaches, and values found in a small region of interest (ROI) located in the grey matter (GM) were in line with previous measurements (T2MRF=88{+/-}7ms vs T2Ref=89{+/-}11ms, T1MRF=1153{+/-}154ms vs T1Ref=1122{+/-}52ms, T2*MRF=56{+/-}4ms vs T2*Ref=53{+/-}3ms).\n\nConclusionAdding a spin echo data segment to EPI based MRF allows accurate and robust measurements of T2, T1 and T2* relaxation times. This MRF framework is easier to implement than spiral-based MRF. It doesnt suffer from undersampling artifacts and seems to require a smaller dictionary size that can fasten the reconstruction process.

biophysics↗

The palmitoyl acyltransferase DHHC5 mediates beta-adrenergic signaling in the heart by targeting Gα proteins and G protein-coupled receptor kinase 2

S-palmitoylation is a reversible posttranslational modification that plays an important role in regulating protein localization, trafficking, and stability. Recent studies have shown that some proteins undergo extremely rapid palmitoylation/depalmitoylation cycles after cellular stimulation supporting a direct signaling role for this posttranslational modification. Here we investigated whether {beta}-adrenergic stimulation of cardiomyocytes led to stimulus-dependent palmitoylation of downstream signaling proteins. We found that {beta}-adrenergic stimulation led to increased Gs and Gi palmitoylation. The kinetics of palmitoylation was temporally consistent with the downstream production of cAMP and positive inotropic responses. Additionally, we identified for the first time that G protein-coupled receptor kinase 2 (GRK2) is a palmitoylated protein in cardiomyocytes. The kinetics of GRK2 palmitoylation were distinct from those observed with Gs and Gi after {beta}-adrenergic stimulation. Knockdown of the plasma membrane-localized palmitoyl acyltransferase DHHC5 revealed that this enzyme is necessary for palmitoylation of Gs, Gi, and GRK2 and functional responses downstream of {beta}-adrenergic stimulation. Our results reveal that DHHC5 activity is required for signaling downstream of {beta}-adrenergic receptors.

cell biology↗

The anthelmintic niclosamide is a potent TMEM16A antagonist that fully bronchodilates airways

There is an unmet need in severe asthma where approximately 40% of patients exhibit poor {beta}-agonist responsiveness, suffer daily symptoms and show frequent exacerbations. Antagonists of the Ca2+-activated-Cl- channel, TMEM16A, offers a new mechanism to bronchodilate airways and block the multiple contractiles operating in severe disease. To identify TMEM16A antagonists we screened a library of ~580,000 compounds. The anthelmintics niclosamide, nitazoxanide and related compounds were identified as potent TMEM16A antagonists that blocked airway smooth muscle depolarization and contraction. To evaluate whether TMEM16A antagonists resist use- and inflammatory-desensitization pathways limiting {beta}-agonist action, we tested their efficacy under harsh conditions using maximally contracted airways or airways pretreated with a cytokine cocktail. Stunningly, TMEM16A antagonists fully bronchodilated airways, while the {beta}-agonist isoproterenol showed only partial effects. Thus, antagonists of TMEM16A and repositioning of niclosamide and nitazoxanide represent an important additional treatment for patients with severe asthma and COPD that is poorly controlled with existing therapies. It is of note that drug repurposing has also attracted wide interest in niclosamide and nitazoxanide as a new treatment for cancer and infectious disease. For the first time we identify TMEM16A as a molecular target for these drugs and thus provide fresh insights into their mechanism for the treatment of these disorders in addition to respiratory disease.

physiology↗

Tau Internalization is Regulated by 6-O Sulfation on Heparan Sulfate Proteoglycans (HSPGs)

The misfolding and accumulation of tau protein into intracellular aggregates known as neurofibrillary tangles is a pathological hallmark of neurodegenerative diseases such as Alzheimers disease. However, while tau propagation is a known marker for disease progression, exactly how tau propagates from one cell to another and what mechanisms govern this spread are still unclear. Here, we report that cellular internalization of tau is regulated by quaternary structure and have developed a cellular assay to screen for genetic modulators of tau uptake. Using CRISPRi technology we have tested 3200 genes for their ability to regulate tau entry and identified enzymes in the heparan sulfate proteoglycan biosynthetic pathway as key regulators. We show that 6-O-sulfation is critical for tauheparan sulfate interactions and that this modification regulates uptake in human central nervous system cell lines, iPS-derived neurons, and mouse organotypic brain slice culture. Together, these results suggest novel strategies to halt tau transmission.

neuroscience↗