Biology subjects

Hara, M.

Publications and source records attributed to Hara, M..

At least 19 recordsLinked to original sources

bioRxiv2026

Islet size and spatial organization govern endocrine cell composition throughout the human pancreas

Stem cell-derived (SC) islets hold promise as a transformative therapy for type 1 diabetes (T1D). To enhance graft uniformity, SC-islet organoids are often dissociated and reaggregated into standardized sizes. However, how endocrine cell composition varies with islet size in native human islets remains largely unknown. Defining this relationship is essential for establishing physiological benchmarks for islet engineering for transplantation. In this study, we examined the relationships among islet size, endocrine cell composition, and spatial distribution throughout whole pancreata from 10 age-matched, non-diabetic male subjects. Insulin-, glucagon-, and somatostatin-positive endocrine cells were identified as single-, double-, and triple-hormone populations ranging from non-islet single endocrine cells and small clusters (niSSCs) to large islets. Approximately 80% of the total endocrine area was contained in islets positive for all three hormones. Islet size strongly correlated with endocrine composition. Specifically, larger islets had increased alpha-cell fractions and reduced beta-cell fractions, while delta-cell fractions were consistent across sizes. Spatial analysis revealed that niSSCs, mono-, and tri-hormone islets frequently exhibit a clustered distribution. Additionally, tri-hormone islets were positioned near the core of the pancreas, whereas mono-somatostatin islets were often localized in the periphery. These findings were consistent across all three pancreatic regions (head, body and tail).

pathology↗

bioRxiv2026

Myeloid STING restrains cardiac remodeling by suppressing macrophage amyloid precursor protein

Mitochondrial DNA (mtDNA) released into the cytosol activates innate immune signaling and promotes inflammation, yet its role in macrophages following sterile tissue injury remains poorly understood. Here, we show that cardiac macrophages from both patients and mice with myocardial infarction (MI) exhibit increased mitochondrial biogenesis, mitochondrial content, membrane potential, and expression of mitochondrial nucleases that facilitate mtDNA release. Consistently, macrophage-specific silencing of genes regulating mitochondrial biogenesis or mtDNA processing attenuated adverse cardiac remodeling after MI. Unexpectedly, despite the role of mtDNA in activating the cGAS-STING pathway, myeloid deletion or macrophage-specific silencing of Sting or cGas exacerbated ventricular dilation, fibrosis, and contractile dysfunction following MI. Single-cell transcriptomic and cell communication analyses identified amyloid precursor protein (APP) as a key downstream effector of STING in cardiac macrophages. Macrophage-specific in vivo App silencing rescued the detrimental effects of myeloid Sting deficiency, establishing APP as a critical mediator of adverse remodeling. Mechanistically, STING interacted with the transcriptional repressor MZF1, promoted its nuclear localization, facilitated its binding to the App promoter, and suppressed App transcription to restrain adverse cardiac remodeling. Together, our findings uncover an unexpected cardioprotective function of myeloid STING and identify the STING-MZF1-APP axis as a previously unrecognized mechanism governing cardiac repair after myocardial infarction.

cell biology↗

bioRxiv2026

Food preference is associated with distinct large-scale cortical functional connectivity patterns during food-image observation

Food preference influences behavior toward food-related stimuli, yet the large-scale neural mechanisms underlying this process remain unclear. This study investigated whether preferred and nonpreferred food cues are associated with distinct patterns of cortical functional connectivity during the observation of food images. Data from 25 of the 40 recruited healthy adults were included in the final analysis after excluding individuals with highly unbalanced response tendencies. Participants viewed 150 food images and rated each image on a four-point preference scale. Trials were classified as favorite food (FF) or disliked food (DF). High-density electroencephalography (EEG) was recorded during the task, and source-level ROI-to-ROI functional connectivity was analyzed using amplitude envelope correlation in the alpha (8-13 Hz) and beta (13-25 Hz) frequency bands over the 1000-ms period after food-picture onset. Response time did not differ significantly between FF and DF trials. However, distinct functional connectivity patterns were observed between conditions in both frequency bands. In the alpha band, FF trials involved a network including the cuneus, parietal regions, cingulate regions, and lateral occipital cortex, whereas DF trials involved the isthmus cingulate, caudal middle frontal gyrus, inferior temporal cortex, superior parietal lobule, and lateral occipital cortex. In the beta band, FF trials involved the isthmus cingulate, precuneus, parietal regions, and pericalcarine cortex, whereas DF trials additionally involved frontal regions, including the superior frontal gyrus and pars triangularis. These findings indicate that food preference is associated with distinct large-scale cortical functional connectivity patterns during food image observation, suggesting differential neural processing of preferred and nonpreferred food cues.

neuroscience↗

bioRxiv2025

Presence Hallucination Induction through Robotically Mediated Somatomotor Conflicts: a pooled analysis of 26 experiments

Hallucinations are significant symptoms in psychiatric and neurodegenerative diseases, that may indicate advanced disease progression or worse disease forms. They are also frequent in healthy individuals, especially elderly or bereaved. Despite their relevance, studying hallucinations in controlled laboratory conditions remains challenging given the limited procedures inducing clinically relevant hallucinations. We have previously developed a robotics-based protocol capable of inducing a specific clinically relevant hallucination, presence hallucination (PH), in both healthy individuals and patients. Using this approach, we have systematically investigated the sensitivity and intensity of PH-induction, as well as its effects on various sensory, behavioral and cognitive aspects. In the present study, we pooled individual-participant data from 26 in-house experiments (totaling 580 individuals) and conducted a Bayesian analysis to estimate effects and moderators of PH-induction. PH-induction was reliably induced with a medium effect-size, and individuals with schizotypal traits were more sensitive. Furthermore, we identified that PH-induction may not strongly depend on altered agency, but found a synergistic relationship between passivity and induced PH. Collectively these results elucidate the role of somatomotor processes in aberrant own body perceptions, advance understanding of psychosis, and provide powerful statistical priors for future studies.

neuroscience↗

bioRxiv2025

Functional genetics with a hypomorphic CENP-C mutant reveals a regulatory system for chromosome congression

Chromosome congression is a key process that acts to align chromosomes at the spindle equator via kinetochore-microtubule interactions, with defects in chromosome alignment leading to chromosomal instability. However, defining the mechanisms that underlie chromosome congression is limited due to the multiple factors that act in parallel to regulate chromosome movement. Here, we conducted a genome-wide Cas9-based functional genetics screen using a hypomorphic CENP-C mutant that affects its kinetochore interactions. Our analysis identified KIF18A, whose knockout resulted in synthetic lethality with the CENP-C mutant. Further analysis revealed that the synthetic defect was due to a reduction in CENP-E function in the CENP-C mutant. Our work suggests that KIF18A promotes chromosome alignment in cooperation with CENP-E downstream of CENP-C during early prometaphase. Thus, our analysis enables us to dissect parallel molecular mechanisms for chromosome congression and identify sensitivities and biomarkers that might guide anti-KIF18A chemotherapeutics.

cell biology↗

bioRxiv2025

High-Throughput Measurement of Oxidative Stress Indicators: d-ROMs and BAP Assays in a 384-Well Plate Format

The bodys oxidative balance is regulated by systems that produce and eliminate reactive oxygen species (ROS), and disruptions in this balance can contribute to the development of lifestyle-related diseases. d-ROMs and BAP tests, which evaluate oxidative balance using just 30 {micro}L of serum per sample in approximately 10 minutes, are widely used for this purpose. However, these methods are not suitable for large-scale studies due to their low-throughput and cost. To overcome these limitations, we established a simultaneous measurement system for multiple samples (144 samples) for measuring d-ROMs and BAP using a pipetting robot and a 384-well plate. The developed system demonstrated good linearity and reproducibility, while significantly reducing reagent and sample consumption. Measurement differences compared to the one of the currently available FREE Carrio Duo system ranged from -5% to 3% for d-ROMs and -12% to 8% for BAP, indicating high consistency with existing methods. Furthermore, the measurement time was substantially shortened from 24 hours (10 minutes x 144 samples) to at longest four hours, although the final color measurements for d-ROMs were performed one week later. This optimized semi-automated system enables the precise and efficient measurement of oxidative stress markers, making it suitable for large-scale studies.

biochemistry↗

bioRxiv2025

The role of GATA2 in the expression of the soluble decoy receptor ST2/IL1RL1 in human and mouse mast cells

The ST2/IL1RL1 gene encodes a receptor subunit for IL-33. The ST2/IL1RL1 gene is transcribed and translated to two splice variants, full-length ST2 (ST2L) and soluble ST2 (sST2). ST2L is expressed on cell surface as membrane bound molecule and forms a heterodimer with IL-1RAcP, which plays an important role in Th2 transduces a signaling of IL-33. In contrast, sST2 blocks the IL-33-signaling via functioning as a decoy receptor of IL-33. In this study, we analyzed the regulatory mechanisms of the gene expression of sST2 in mast cell, the major source of sST2. We found that a hematopoietic cell-specific transcription factor GATA2 is essential for sST2 expression in human mast cells (a cell line and primary cells) and mouse mast cells. Various assays including ChIP assay, reporter assay, and 3C assay revealed that GATA2 binding to the distal promoter of the IL1RL1 gene transcriptionally activates proximal promoter-driven expression of sST2 through chromosomal conformation.

immunology↗

bioRxiv2024

Identification of the sex determination region and the development of a marker to distinguish males and females in megai abalone (Haliotis gigantea)

Sex identification markers are of considerable value to aquaculture and fisheries as they enable pre-maturity prediction of the genotypic sex of individuals as well as of the sex ratio in offspring. For these reasons, extensive investigations of the mechanisms of sex determination have been conducted in commercially important finfish species, resulting in the development of sex identification markers for many of these. However, research on shellfish is still in its infancy stage. In this study, we investigated the genomic basis of sex determination in megai abalone (Haliotis gigantea), a gastropod species of economic importance in fisheries and aquaculture in East Asia, with the aim of developing a sex identification marker. Initially, we examined the applicability of sex identification markers reported for two other Haliotis species, but found they were not transferable to H. gigantea. We then analyzed the sex chromosomes in H. gigantea using a whole-genome resequencing approach. By calculating the fixation index value between 10 females and 10 males, we identified a genomic region spanning approximately 2 Mb that exhibited nucleotide sequence divergence between the sexes (from the position 260,372 bp to 1,930,278 bp in chromosome 18). This region was located next to the pseudoautosomal region which is characterized by an absence of sequence differentiation between the sexes. In the divergent region, male-specific SNPs were identified but no female-specific SNPs were detected, indicating that this species has an XX-XY sex determination system. By exploring Indel polymorphisms in the divergent region, we successfully developed a codominant PCR marker for sex identification that could be applied to wild individuals from geographically distant locations. A cross-species analysis suggested that the sex determination region in H. gigantea was unlikely to be shared with the closely related species H. discus hannai. This raises the possibility of a widespread, rapid turnover of sex chromosomes or sex determination regions in abalones, similar to certain vertebrate lineages such as finfish.

genomics↗

bioRxiv2024

Molecular details and phospho-regulation of the CENP-T-Mis12 complex interaction during mitosis in DT40 cells

To establish bi-polar attachments of microtubules to sister chromatids, an inner kinetochore subcomplex, the constitutive centromere-associated network (CCAN), is assembled on centromeric chromatin and recruits the microtubule-binding subcomplex called the KMN network. Although it is important to characterize the interaction between CCAN and KMN, it is difficult to evaluate the significance of each interaction in cells, since CCAN proteins CENP-C and CENP-T independently bind to the Mis12 complex (Mis12C) of KMN. Here, we demonstrate molecular details of the CENP-T-Mis12C interaction using chicken DT40 cells lacking CENP-C-Mis12C interaction. Based on AlphaFold2 predictions, we identified two binding surfaces for the CENP-T-Mis12C interaction, demonstrating that each surface is critical for Mis12C recruitment and cell viability. This interaction via two interaction surfaces is cooperatively regulated by dual phosphorylation of Dsn1 (a Mis12C component) and CENP-T, ensuring robust CENP-T-Mis12C interaction and proper mitotic progression. These findings deepen our understanding of kinetochores assembly in cells.

cell biology↗

bioRxiv2024

CENP-C-Mis12 complex establishes a regulatory loop through Aurora B for chromosome segregation

Establishing the correct kinetochore-microtubule attachment is crucial for faithful chromosome segregation. The kinetochore has various regulatory mechanisms for establishing correct bipolar attachment. However, how the regulations are coupled is not fully understood. Here, we demonstrate a regulatory loop between the kinetochore protein CENP-C and Aurora B kinase, which is critical for the error correction of kinetochore-microtubule attachment. This regulatory loop is mediated through the binding of CENP-C to the outer kinetochore Mis12 complex (Mis12C). Although the Mis12C binding region of CENP-C is dispensable for mouse development and proliferation in human RPE-1 cells, those cells lacking this region display increased mitotic defects. The CENP-C-Mis12C interaction facilitates the centromeric recruitment of Aurora B and the mitotic error correction in human cells. Given that Aurora B reinforces the CENP-C-Mis12C interaction, our findings reveal a positive regulatory loop between Aurora B recruitment and the CENP-C-Mis12C interaction, which ensures chromosome bi-orientation for accurate chromosome segregation.

cell biology↗

bioRxiv2023

Microfluidic device for simple diagnosis of plant growth condition by detecting miRNAs from filtered plant extracts

Plants are exposed to a variety of environmental stress and starvation of inorganic phosphorus can be a major constraint in crop production. In plants, in response to phosphate deficiency in soil, miR399, a type of microRNA (miRNA), is upregulated. By detecting miR399, the early diagnosis of phosphorus deficiency stress in plants can be accomplished. However, general miRNA detection methods require complicated experimental manipulations. Therefore, simple and rapid miRNA detection methods are required for early plant nutritional diagnosis. For the simple detection of miR399, microfluidic technology is suitable for point-of-care applications because of its ability to detect target molecules in small amounts in a short time and with simple manipulation. In this study, we developed a microfluidic device to detect miRNAs from filtered plant extracts for the easy diagnosis of plant growth conditions. To fabricate the microfluidic device, verification of the amine-terminated glass as the basis of the device and the DNA probe immobilization method on the glass was conducted. In this device, the target miRNAs were detected by fluorescence of sandwich hybridization in a microfluidic channel. For plant stress diagnostics using a microfluidic device, we developed a protocol for miRNA detection by validating the sample preparation buffer, filtering, and signal amplification. Using this system, endogenous sly-miR399 in tomatoes, which is expressed in response to phosphorus deficiency, was detected before the appearance of stress symptoms. This early diagnosis system of plant growth conditions has a potential to improve food production and sustainability through cultivation management.

plant biology↗

bioRxiv2023

Transfer of cyclobutane pyrimidine dimer photolyase to chloroplasts for Poaceae survival under ultraviolet-B radiation

Cyclobutane pyrimidine dimer (CPD) photolyase (PHR), the primary enzyme for repairing the CPD induced by ultraviolet B (UV-B) radiation, is essential for plants living under sunlight. Rice CPD photolyase (OsPHR), is such a unique triple-targeting protein. The signal sequences required for its translocation to the nucleus or mitochondria are located in the C-terminal region but were yet to be identified for chloroplasts. Here, we identified sequences located in the N-terminal region, including the serine-phosphorylation site at position 7 of OsPHR, and found that OsPHR is transported/localized to chloroplasts via a vesicle transport system under the control of serine phosphorylation. However, the sequence identified in this study is only conserved in some Poaceae species and in many other plants, PHR does not localize to chloroplasts Therefore, we reasoned that Poaceae species need the ability to repair CPD in the chloroplast genome to survive under sunlight and have acquired this new mechanism for chloroplast translocation.

cell biology↗

bioRxiv2022

High-resolution structure of a microtubule-like tube composed of FtsZ-monobody complexes

FtsZ, a bacterial tubulin homologue, forms protofilaments and the Z-ring, which acts as a scaffold for accessory proteins during cell division. Although various studies have revealed its molecular mechanisms, the lack of high-resolution solution structures has hindered the understanding of the detailed mechanisms. Here, we developed a monobody (Mb) that binds FtsZs from Escherichia coli and Klebsiella pneumoniae (KpFtsZ) without affecting their GTPase activities. When expressed in E. coli cells, the Mb did not inhibit Z-ring formation but did inhibit cell division. The crystal structures of the KpFtsZ-Mb complexes revealed the epitope, and the cryoEM structure at 2.67 [A] resolution showed a double helical tube consisting of two KpFtsZ protofilaments stabilized by the Mb filling interfilament gaps. Our structural analyses highlight the similarity between the microtubule and the FtsZ tube and the importance of the plasticity of FtsZ protofilaments.

molecular biology↗

bioRxiv2022

The centromere/kinetochore is assembled through CENP-C oligomerization

The kinetochore is an essential protein complex for accurate chromosome segregation. The constitutive centromere-associated network (CCAN), a subcomplex of the kinetochore, associates with centromeric chromatin providing a platform for the kinetochore assembly. A CCAN protein, CENP-C, is thought to be a central hub for the centromere/kinetochore organization. However, the crucial role of CENP-C in centromeres remains to be elucidated. Here, we demonstrated that both the CCAN-binding domain and C-terminal Cupin domain of CENP-C are necessary and sufficient for chicken CENP-C function. Our structural and biochemical analyses revealed that the Cupin domain of chicken and human CENP-C is self-oligomerization domain, which is crucial for centromeric chromatin organization. CENP-C mutants lacking the oligomerization interface cause mislocalization of CCAN and cell death. Based on these results, we conclude that the CENP-C oligomerization plays a crucial role in centromere function via providing the robust centromeric chromatin in vertebrate cells.

cell biology↗

bioRxiv2021

Identification of CD73 as the antigen of an antigen-unknown monoclonal antibody established by exosome immunization, and its antibody-drug conjugate exerts an antitumor effect on glioblastoma cell lines

Development of antibodies against the native structure of membrane proteins with multiple transmembrane domains is challenging because it is difficult to prepare antigens with native structures. Previously, we successfully developed a monoclonal antibody against multi-pass membrane protein TMEM180 by exosome immunization in rats. This approach yielded antibod-ies that recognized cancer-specific antigens on the exosome. In this study, we performed im-munoprecipitation using magnetic beads to identify the antigen of one of the rat antibody clones, 0614, as CD73. We then converted antibody 0614 to human chimeric antibody 0614-5 and found that the humanized antibody did not inhibit CD73 enzymatic activity. Glioblastoma (GB) was the cancer type with the highest expression of CD73 in the tumor relative to healthy tissue. An antibody-drug conjugate (ADC) of 0614-5 exerted an antitumor effect on GB cell lines according to expression of CD73. The 0614-5-ADC could be used to treat cancers with high CD73 expression. In addition, our strategy could be used to determine the antigen of any antibody produced by exosome immunization.

biophysics↗

bioRxiv2021

Tactile spatial discrimination on the torso using vibrotactile and force stimulation

There is a steadily growing number of mobile communication systems that provide spatially encoded tactile information to the humans torso. However, the increased use of such hands-off displays is currently not matched with or supported by systematic perceptual characterization of tactile spatial discrimination on the torso. Furthermore, there are currently no data testing spatial discrimination for dynamic force stimuli applied to the torso. In the present study, we measured tactile point localization (PL) and tactile direction discrimination (DD) on the thoracic spine using two unisex torso-worn tactile vests realized with arrays of 3x3 vibrotactile or force feedback actuators. We aimed to, firstly, evaluate and compare the spatial discrimination of vibrotactile and force stimulations on the thoracic spine and, secondly, to investigate the relationship between the PL and DD results across stimulations. Thirty-four healthy participants performed both tasks with both vests. Tactile accuracies for vibrotactile and force stimulations were 60.7% and 54.6% for the PL task; 71.0% and 67.7% for the DD task, respectively. Performance correlated positively with both stimulations, although accuracies were higher for the vibrotactile than for the force stimulation across tasks, arguably due to specific properties of vibrotactile stimulations. We observed comparable directional anisotropies in the PL results for both stimulations; however, anisotropies in the DD task were only observed with vibrotactile stimulations. We discuss our findings with respect to tactile perception research as well as their implications for the design of high-resolution torso-mounted tactile displays for spatial cueing.

neuroscience↗

bioRxiv2020

Cogno-Vest: A Torso-Worn, Force Display to Experimentally Induce Specific Hallucinations and Related Bodily Sensations

Recent advances in virtual reality and robotic technologies have allowed researchers to explore the mechanisms underlying bodily aspects of self-consciousness which are largely attributed to the multisensory and sensorimotor processing of bodily signals (bodily self-consciousness, BSC). One key contribution to BSC, that is currently poorly addressed due to the lack of a wearable solution, concerns realistic collision sensations on the torso. Here, we introduce and validate a novel torso-worn force display, the Cogno-vest, to provide mechanical touch on the users back in a sensorimotor perception experiment. In a first empirical study, we characterized human finger poking (N=28). In order to match these poking characteristics and meet the wearability criteria, we used bi-directional, push-pull solenoids as a force actuator in the Cogno-vest. Subsequently, and based on an iterative, multidisciplinary design procedure, a bodyconforming, unisex, torso-worn force display was prototyped. Finally, we conducted a behavioral study that investigated BSC in 25 healthy participants by introducing conflicting sensorimotor signals between their hand and torso (back). Using the final reiteration of the Cogno-vest we successfully replicated previous findings on illusory states of BSC, characterized by presence hallucinations (PH) and passivity symptoms, and achieved higher illusion ratings compared to static conditions used in prior studies.

neuroscience↗

bioRxiv2020

Sensorimotor hallucinationsin Parkinson's disease

Hallucinations in Parkinsons disease (PD) are one of the most disturbing non-motor symptoms, affect half of the patients, and constitute a major risk factor for adverse clinical outcomes such as psychosis and dementia. Here we report a robotics-based approach, enabling the induction of a specific clinically-relevant hallucination (presence hallucination, PH) under controlled experimental conditions and the characterization of a PD subgroup with enhanced sensorimotor sensitivity for such robot-induced PH. Using MR-compatible robotics in healthy participants and lesion network mapping analysis in neurological non-PD patients, we identify a fronto-temporal network that was associated with PH. This common PH-network was selectively disrupted in a new and independent sample of PD patients and predicted the presence of symptomatic PH. These robotics-neuroimaging findings determine the behavioral and neural mechanisms of PH and reveal pathological cortical sensorimotor processes of PH in PD, identifying a more severe form of PD associated with psychosis and cognitive decline.

neuroscience↗