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Lee, M.-H.

Publications and source records attributed to Lee, M.-H..

30 records · Page 2Linked to original sources

Multi-compartment tumor organoids

Organoid cultures are widely used because they preserve many features of cancer cells in vivo. Here, we developed high-throughput oil-in-water droplet microtechnology to generate highly uniform, small-volume, multi-compartment organoids. Each organoid culture features a microenvironmental architecture that mimics both the basement membrane and stromal barriers. This matrix architecture, which allows accessing both proliferative and invasive features of cancer cells in a single platform, has profound effect on observed drug responsiveness and tumor progression that correlate well with in vivo and clinical outcomes. The method was tested on multiple types of cancer cells including primary cells and immortalized cell lines, and we determined our platform is suitable even for cells of poor organoid-forming ability. These new organoids also allow for direct orthotopic mouse implantation of cancer cells with unprecedented success.

cancer biology↗

E-cadherin promotes cell hyper-proliferation in breast cancer

The loss of the intercellular adhesion molecule E-cadherin is a hallmark of the epithelial- mesenchymal transition (EMT), during which tumor cells transition into an invasive phenotype. Accordingly, E-cadherin has long been considered a tumor suppressor gene. Using novel multi-compartment spheroids and multiple in vivo models, we show that E-cadherin promotes a hyper-proliferative phenotype in breast cancer cells via interaction with the transmembrane receptor EGFR. This interaction results in the activation of the MEK/ERK signaling pathway, leading to a significant increase in proliferation via the activation of transcription factors including c-Fos. Pharmacological inhibition of MEK activity in E-cadherin positive breast cancer cells significantly decreases both tumor growth and macro-metastasis in vivo. This work provides evidence for a novel role of E-cadherin in breast tumor growth and identifies a potential new target to treat hyper-proliferative E-cadherin-positive breast tumors.

cancer biology↗

IFN-{lambda}4 may contribute to HCV persistence by increasing ER stress and enhancing IRF1 signaling

Chronic hepatitis C virus (HCV) infection and cirrhosis are major risk factors for developing hepatocellular carcinoma (HCC). Genetic polymorphisms in the IFNL3/IFNL4 locus have been associated both with impaired clearance of HCV and protection from liver fibrosis, an early stage of cirrhosis. Here, we aimed to address the genetic and functional relationships between IFNL3/IFNL4 polymorphisms, HCV-related cirrhosis, and HCC risk. We evaluated associations between IFNL4 genotype, defined as the presence of rs368234815-dG or rs12979860-T alleles, with cirrhosis and HCC risk in patients with chronic HCV - 2,931 from Taiwan and 3,566 from Japan. We detected associations between IFNL4 genotype and decreased risk of cirrhosis (OR=0.66, 95%CI=0.46-0.93, P=0.018, in Taiwan), but increased risk of HCC (OR=1.28, 95%CI=1.07-1.52, P=0.0058, in Japan). In-vitro, IFN-{lambda}4 expression increased ER stress, and enhanced positive regulation of IFN responses via IRF1 induction, which mediated antiproliferative effects in hepatic cells. Our data present novel IFN-{lambda}4-associated pathways that may be contributing to HCV persistence and development of HCC.

cancer biology↗

Motor variability prior to learning is a poor predictor of the ability to adopt new movement solutions

Many contexts in motor learning require a learner to change from an existing movement solution to a novel movement solution to perform the same task. Recent evidence has pointed to motor variability prior to learning as a potential marker for predicting individual differences in motor learning. However, it is not known if this variability is predictive of the ability to adopt a new movement solution for the same task. Here, we examined this question in the context of a redundant precision task requiring control of motor variability. Fifty young adults learned a precision task that involved throwing a virtual puck toward a target using both hands. Because the speed of the puck depended on the sum of speeds of both hands, this task could be achieved using multiple solutions. Participants initially performed a baseline task where there was no constraint on the movement solution, and then performed a novel task where they were constrained to adopt a specific movement solution requiring asymmetric left and right hand speeds. Results showed that participants were able to learn the new solution, and this change was associated with changes in both the amount and structure of variability. However, individual differences in baseline motor variability were only weakly correlated with initial and final task performance when using the new solution, with greater variability being associated with higher errors. We also found a strong specificity component - initial variability when using the new solution was highly correlated with final task performance with the new solution, but once again, higher variability was associated with greater errors. These results suggest that motor variability is not necessarily indicative of flexibility and highlight the need to consider the task context in determining the relation between motor variability and learning.

neuroscience↗

No evidence of coronaviruses or other potentially zoonotic viruses in Sunda pangolins (Manis javanica) entering the wildlife trade via Malaysia.

The legal and illegal trade in wildlife for food, medicine and other products is a globally significant threat to biodiversity that is also responsible for the emergence of pathogens that threaten human and livestock health and our global economy. Trade in wildlife likely played a role in the origin of COVID-19, and viruses closely related to SARS-CoV-2 have been identified in bats and pangolins, both traded widely. To investigate the possible role of pangolins as a source of potential zoonoses, we collected throat and rectal swabs from 334 Sunda pangolins (Manis javanica) confiscated in Peninsular Malaysia and Sabah between August 2009 and March 2019. Total nucleic acid was extracted for viral molecular screening using conventional PCR protocols used to routinely identify known and novel viruses in extensive prior sampling (>50,000 mammals). No sample yielded a positive PCR result for any of the targeted viral families - Coronaviridae, Filoviridae, Flaviviridae, Orthomyxoviridae and Paramyxoviridae. In light of recent reports of coronaviruses including a SARS-CoV-2 related virus in Sunda pangolins in China, the lack of any coronavirus detection in our upstream market chain samples suggests that these detections in downstream animals more plausibly reflect exposure to infected humans, wildlife or other animals within the wildlife trade network. While confirmatory serologic studies are needed, it is likely that Sunda pangolins are incidental hosts of coronaviruses. Our findings further support the importance of ending the trade in wildlife globally.

molecular biology↗

Controlling a robotic arm for functional tasks using a wireless head-joystick: A case study of a child with congenital absence of upper and lower limbs

Children with movement impairments needing assistive devices for activities of daily living often require novel methods for controlling these devices. Body-machine interfaces, which rely on body movements, are particularly well-suited for children as they are non-invasive and have high signal-to-noise ratios. Here, we examined the use of a head-joystick to enable a child with congenital absence of all four limbs to control a seven degree-of-freedom robotic arm. Head movements were measured with a wireless inertial measurement unit and used to control a robotic arm to perform two functional tasks - a drinking task and a block stacking task. The child practiced these tasks over multiple sessions; a control participant performed the same tasks with a manual joystick. Our results showed that the child was able to successfully perform both tasks, with movement times decreasing by ~40-50% over 6-8 sessions of training. The childs performance with the head-joystick was also comparable to the control participant using a manual joystick. These results demonstrate the potential of using head movements for the control of high degree-of-freedom tasks in children with limited movement repertoire.

neuroscience↗

Self-controlled practice and nudging during structural learning of a novel control interface

Self-controlled practice schedules have been shown to enhance motor learning in several contexts, but their effectiveness in structural learning tasks, where the goal is to eventually learn an underlying structure or rule, is not well known. Here we examined the use of self-controlled practice in a novel control interface requiring structural learning. In addition, we examined the effect of nudging - i.e., whether altering task difficulty could influence self-selected strategies, and hence facilitate learning. Participants wore four inertial measurement units (IMUs) on their upper body and the goal was to use motions of the upper body to move a screen cursor to different targets presented on the screen. The structure in this task that had to be learned was based on the fact that the signals from the IMUs were linearly mapped to the x- and y-position of the cursor. Participants (N = 62) were split into 3 groups (random, self-selected, nudge) based on whether they had control over the sequence in which they could practice the targets. To test whether participants learned the underlying structure, participants were tested both on the trained targets, as well as novel targets that were not practiced during training. Results showed that during training, the self-selected group showed shorter movement times relative to the random group, and both self-selected and nudge groups adopted a strategy of tending to repeat targets. However, in the test phase, we found no significant differences in task performance between groups, indicating that structural learning was not reliably affected by the type of practice. In addition, nudging participants by adjusting task difficulty did not show any significant benefits to overall learning. These results suggest that although self-controlled practice influenced practice structure and facilitated learning, it did not provide any additional benefits relative to practicing on a random schedule in this task.

neuroscience↗

Reply: Evidence that APP gene copy number changes reflect recombinant vector contamination

In the accompanying comment1, Kim et al. concluded that somatic gene recombination (SGR) and amyloid precursor protein (APP) genomic complementary DNAs (gencDNAs) in brain are contamination artifacts and do not naturally exist. We disagree. Here we address the three types of analyses used by Kim et al. to reach their conclusions: informatic contaminant identification, plasmid PCR, and single-cell sequencing. Additionally, Kim et al. requested "reads supporting novel APP insertion breakpoints," and we now provide 10 different examples that support APP gencDNA insertion within eight chromosomes beyond wildtype APP on chromosome 21 from Alzheimers disease (AD) samples. If SGR exists as experimentally supported here and previously2,3, contamination scenarios become moot. Our informatic analyses of data generated by an independent laboratory (Park et al.)4, complement and are entirely consistent with what Lee et al.2 presented via nine distinct lines of evidence, in addition to three from a prior publication3. Plasmid contamination was identified in a single pull-down dataset after publication of Lee et al.2; however subsequent analyses did not alter any of our conclusions including those of our prior publications3,5 and plasmid contamination-free replication of this approach by ourselves and others supported the original conclusions. Novel retro-insertion sites, alterations of APP gencDNA number and form with cell type from the same brain and pathogenic SNVs occurring only in AD, all support the existence of APP gencDNAs produced by SGR.

genomics↗

Conformational sensors and domain-swapping reveal structural and functional differences between β-arrestin isoforms

Desensitization, signaling and trafficking of G protein-coupled receptors (GPCRs) are critically regulated by multifunctional adaptor proteins, {beta}-arrestins ({beta}arrs). The two isoforms of {beta}arrs ({beta}arr1 and 2) share a high degree of sequence and structural similarity, still however, they often mediate distinct functional outcomes in the context of GPCR signaling and regulation. A mechanistic basis for such a functional divergence of {beta}arr isoforms is still lacking. Using a set of complementary approaches including antibody fragment based conformational sensors, we discover structural differences between {beta}arr1 and 2 upon their interaction with activated and phosphorylated receptors. Interestingly, domain swapped chimeras of {beta}arrs display robust complementation in functional assays thereby, linking the structural differences between the receptor-bound {beta}arr1 and 2 with their divergent functional outcomes. Our findings reveal important insights into the ability of {beta}arr isoforms to drive distinct functional outcomes, and underscore the importance of integrating this aspect in the current framework of biased agonism.

biophysics↗

Within-finger maps of tactile and nociceptive input in the human parietal cortex

The spatial representation of nociceptive input in the human parietal cortex is not fully understood. For instance, it is known that the primary somatosensory cortex (S1) contains a representation of nociceptive-selective input to different fingers, but it is unclear whether S1 subdivisions contain finer-grained, within-finger maps of nociceptive input. It is also unknown whether within-finger maps of somatosensory input exist in intraparietal regions. Therefore, we used high-field 7T functional MRI to reveal within-finger maps of nociceptive and tactile inputs in the human parietal cortex. Although we replicated the previous findings of between-finger maps of nociceptive input spanning S1 subdivisions, we found weak and inconsistent evidence for within-finger maps of nociceptive input in S1 subdivisions. In the same subjects, we found mirrored, within-finger maps of tactile input in areas 3a, 3b, and 1. Importantly, we discovered a within-finger map of nociceptive input, but not of tactile input, in the human intraparietal area 1 (hIP1). In conclusion, our study indicates that the spatial representation of nociceptive input in the parietal cortex partly differs from that of tactile input and reports the discovery of a within-finger map of nociceptive input in hIP1.\n\nNew & NoteworthyWe report the discovery of a fine-grained map of nociceptive input to finger segments in the human intraparietal area 1.

neuroscience↗

HBx enhances CPAP expression via interacting with CREB to promote hepatocarcinogenesis in HBV-associated HCC

Hepatitis B virus (HBV) encoded non-structure protein X (HBx) can promote cell proliferation, migration, and anti-apoptosis via activating several transcription factors and increasing their downstream gene expression in HBV-infected liver cells. Our previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for HBx-mediated NF-{kappa}B activation. Here, we found that, upon HBV infection, overexpressed HBx can transcriptionally up-regulate CPAP via interacting with CREB. CPAP can directly interact with HBx to promote HBx-mediated cell proliferation and migration; and SUMO modification of CPAP is involved in interacting with HBx. Interestingly, CPAP can increase the HBx protein stability in an NF-{kappa}B-dependent manner; and overexpressed CPAP and HBx is positively correlated with the activation status of NF-{kappa}B in HCC. Increased expression of CREB and CPAP mRNAs exists in the high-risk group with a lower survival rate in hepatocellular carcinoma (HCC). These results suggest that the reciprocal regulation between CPAP and HBx may provide a microenvironment to facilitate HCC development via enhancing NF-{kappa}B activation, inflammatory cytokine production, and cancer maligancies. The findings of this study not only shed light on the role of CPAP in HBV-associated HCC, but also provide CPAP as a potential target for HBV-related HCC therapy.\n\nAuthor SummaryIn this study, we address a novel molecular mechanism for the collaboration between overexpressed HBx and CPAP in promoting hepatocarcinogenesis in HBV-associated HCC. Upon HBV infection, HBx is overexpressed and interacts with CREB to transcriptionally activate CPAP; the HBx/CPAP interaction promotes hepatocarcinogenesis. Clinical analysis found that co-overexpressed CPAP and CREB exist in the high-risk group with a lower survival rate in HCC. Additionally, overexpressed CPAP contributes to HBx protein stability in a NF-{kappa}B-dependent pathway. Our study provides a potential translational application in targeting CREB-CPAP axis in HBV-associated HCC.

cancer biology↗

DIV-1/PolA2 Promotes GLP-1/Notch-Mediated Cellular Events in Caenorhabditis elegans

Notch signaling is a highly conserved cell signaling system in most multicellular organisms and plays a critical role in animal development. In various tumor cells, Notch signaling is elevated and has been considered as an important target in cancer treatments. In C. elegans, GLP-1 (one of two C. elegans Notch receptors) activity is required for cell fate specification in germline and somatic tissues. In this study, we have identified div-1 gene as a positive regulator for GLP-1/Notch-mediated cellular events. C. elegans div-1 encodes the B subunit of the DNA polymerase alpha-primase complex and is highly expressed in proliferative germ cells. Functional analyses demonstrated that i) DIV-1 is required for the robust proliferation typical of the germline, ii) loss of DIV-1 enhances and suppresses specific phenotypes that are associated with reduced and elevated GLP-1/Notch activity in germline and somatic tissues, and iii) DIV-1 works together with FBF/PUF proteins, downstream regulators of GLP-1/Notch signaling, to promote germline stem cell (GSC) maintenance and germline proliferation. To maintain GSCs and proliferative cell fate, GLP-1/Notch activity must remain above a threshold for proliferation/differentiation decision. Our results propose that DIV-1 may control the level of threshold for GLP-1/Notch-mediated germline proliferation. PolA2, a mammalian homolog of the C. elegans DIV-1, has been emerged as a therapeutic target for non-small cell lung cancer (NSCLC). Notably, Notch signaling is altered in approximately one third of NSCLCs. Therefore, the discovery of the DIV-1 effect on GLP-1/Notch-mediated cellular events has implications for our understanding of vertebrate PolA2 protein and its influence on stem cell maintenance and tumorigenesis.

developmental biology↗