Notch transcriptional target tmtc1 maintains vascular homeostasis.

Proper lung function requires the maintenance of a tight endothelial barrier while simultaneously permitting the exchange of macromolecules and fluids to underlying tissue. Disruption of this barrier results in an increased vascular permeability in the lungs, leading to acute lung injury. In this study, we set out to determine whether transcriptional targets of Notch signaling function to preserve vascular integrity. We tested the in vivo requirement for Notch transcriptional signaling in maintaining the pulmonary endothelial barrier by using two complementary endothelial-specific Notch loss-of-function murine transgenic models. Notch signaling was blocked using endothelial-specific activation of an inhibitor of Notch transcriptional activation, Dominant Negative Mastermindlike (DNMAML; CDH5CreERT2), or endothelial-specific loss of Notch1 (Notch1f/f; CDH5CreERT2). Both Notch mutants increased vascular permeability with pan-Notch inhibition by DNMAML showing a more severe phenotype in the lungs and in purified endothelial cells. RNA sequencing of primary lung endothelial cells (ECs) identified novel Notch targets, one of which was transmembrane O-mannosyltransferase targeting cadherins 1 (tmtc1). We show that tmtc1 interacts with vascular endothelial cadherin (VE-cadherin) and regulates VE-cadherin egress from the endoplasmic reticulum through direct interaction. Our findings demonstrate that Notch signaling maintains endothelial adherens junctions and vascular homeostasis by a transcriptional mechanism that drives expression of critical factors important for processing and transport of VE-cadherin.

Nanoparticle targeting of de novo profibrotic macrophages mitigates lung fibrosis.

The pathogenesis of lung fibrosis involves hyperactivation of innate and adaptive immune pathways that release inflammatory cytokines and growth factors such as tumor growth factor (TGF)ß1 and induce aberrant extracellular matrix protein production. During the genesis of pulmonary fibrosis, resident alveolar macrophages are replaced by a population of newly arrived monocyte-derived interstitial macrophages that subsequently transition into alveolar macrophages (Mo-AMs). These transitioning cells initiate fibrosis by releasing profibrotic cytokines and remodeling the matrix. Here, we describe a strategy for leveraging the up-regulation of the mannose receptor CD206 in interstitial macrophages and Mo-AM to treat lung fibrosis. We engineered mannosylated albumin nanoparticles, which were found to be internalized by fibrogenic CD206+ monocyte derived macrophages (Mo-Macs). Mannosylated albumin nanoparticles incorporating TGFß1 small-interfering RNA (siRNA) targeted the profibrotic subpopulation of CD206+ macrophages and prevented lung fibrosis. The findings point to the potential utility of mannosylated albumin nanoparticles in delivering TGFß-siRNA into CD206+ profibrotic macrophages as an antilung fibrosis strategy.

Impact of sagittal and coronal pelvic tilt on hip subluxation in non-ambulatory flaccid neuromuscular scoliosis patients following spinal correction.

PURPOSE: To evaluate the actual change in clinical hip pain and hip migration after operation for non-ambulatory flaccid neuromuscular (NM) scoliosis and investigate whether there is an association between hip migration and coronal/sagittal pelvic tilt (CO-PT/SA-PT). PATIENTS AND METHODS: This retrospective, single-center, observational study evaluated a total of 134 patients with non-ambulatory flaccid neuromuscular scoliosis who underwent surgery performed by a single surgeon between 2003 and 2020, with at least 2 years of follow-up period. Operation procedures were conducted in two stages, beginning with L5-S1 anterior release followed by posterior fixation. Radiologic parameters were measured at preoperative, immediate postoperative, and last follow-up periods with clinical hip pain and clinical hip dislocation events. RESULTS: The significant improvements occurred in various parameters after correction surgery for NM scoliosis, containing Cobb's angle of major curve and CO-PT. However, Reimer's hip migration percentage (RMP) was increased on both side of hip (High side, 0.23 ± 0.16 to 0.28 ± 0.21; Low side, 0.20 ± 0.14 to 0.23 ± 0.18). Hip pain and dislocation events were also increased (Visual analog scale score, 2.5 ± 2.3 to 3.6 ± 2.6, P value < 0.05; dislocation, 6-12). Logistic regression analysis of the interactions between ΔRMP(High) and the change of sagittal pelvic tilt (ΔSA-PT) after correction reveals a significant negative association. (95% CI 1.003-1.045, P value = 0.0226). CONCLUSIONS: In cases of non-ambulatory flaccid NM scoliosis, clinical hip pain, and subluxation continued to deteriorate even after correction of CO-PT. There was a relationship between the decrease in SA-PT, and an increase in hip migration percentage on high side, indicating the aggravation of hip subluxation.

Machine learning based estimation of urban on-road CO2 concentration in Seoul.

The urban on-road CO2 emissions will continue to increase, it is therefore essential to manage urban on-road CO2 concentrations for effective urban CO2 mitigation. However, limited observations of on-road CO2 concentrations prevents a full understanding of its variation. Therefore, in this study, a machine learning-based model that predicts on-road CO2 concentration (CO2traffic) was developed for Seoul, South Korea. This model predicts hourly CO2traffic with high precision (R2 = 0.8 and RMSE = 22.9 ppm) by utilizing CO2 observations, traffic volume, traffic speed, and wind speed as the main factors. High spatiotemporal inhomogeneity of hourly CO2traffic over Seoul, with 14.3 ppm by time-of-day and 345.1 ppm by road, was apparent in the CO2traffic data predicted by the model. The large spatiotemporal variability of CO2traffic was related to different road types (major arterial roads, minor arterial roads, and urban highways) and land-use types (residential, commercial, bare ground, and urban vegetation). The cause of the increase in CO2traffic differed by road type, and the diurnal variation of CO2traffic differed according to land-use type. Our results demonstrate that high spatiotemporal on-road CO2 monitoring is required to manage urban on-road CO2 concentrations with high variability. In addition, this study demonstrated that a model using machine learning techniques can be an alternative for monitoring CO2 concentrations on all roads without conducting observations. Applying the machine learning techniques developed in this study to cities around the world with limited observation infrastructure will enable effective urban on-road CO2 emissions management.

Macrophages show higher levels of engulfment after disruption of cis interactions between CD47 and the checkpoint receptor SIRPα.

The macrophage checkpoint receptor SIRPα signals against phagocytosis by binding CD47 expressed on all cells - including macrophages. Here, we found that inhibiting cis interactions between SIRPα and CD47 on the same macrophage increased engulfment ('eating') by approximately the same level as inhibiting trans interactions. Antibody blockade of CD47, as pursued in clinical trials against cancer, was applied separately to human-derived macrophages and to red blood cell (RBC) targets for phagocytosis, and both scenarios produced surprisingly similar increases in RBC engulfment. Blockade of both macrophages and targets resulted in hyper-phagocytosis, and knockdown of macrophage-CD47 likewise increased engulfment of 'foreign' cells and particles, decreased the baseline inhibitory signaling of SIRPα, and linearly increased binding of soluble CD47 in trans, consistent with cis-trans competition. Many cell types express both SIRPα and CD47, including mouse melanoma B16 cells, and CRISPR-mediated deletions modulate B16 phagocytosis, consistent with cis-trans competition. Additionally, soluble SIRPα binding to human CD47 displayed on Chinese hamster ovary (CHO) cells was suppressed by SIRPα co-display, and atomistic computations confirm SIRPα bends and binds CD47 in cis Safety and efficacy profiles for CD47-SIRPα blockade might therefore reflect a disruption of both cis and trans interactions.

Physical and Soluble Cues Enhance Tendon Progenitor Cell Invasion into Injectable Synthetic Hydrogels.

Synthetic hydrogels represent an exciting avenue in the field of regenerative biomaterials given their injectability, orthogonally tunable mechanical properties, and potential for modular inclusion of cellular cues. Separately, recent advances in soluble factor release technology have facilitated control over the soluble milieu in cell microenvironments via tunable microparticles. A composite hydrogel incorporating both of these components can robustly mediate tendon healing following a single injection. Here, a synthetic hydrogel system with encapsulated electrospun fiber segments and a novel microgel-based soluble factor delivery system achieves precise control over topographical and soluble features of an engineered microenvironment, respectively. It is demonstrated that three-dimensional migration of tendon progenitor cells can be enhanced via combined mechanical, topographical, and microparticle-delivered soluble cues in both a tendon progenitor cell spheroid model and an ex vivo murine Achilles tendon model. These results indicate that fiber reinforced hydrogels can drive the recruitment of endogenous progenitor cells relevant to the regeneration of tendon and, likely, a broad range of connective tissues.

Comparison of the efficacy between sequential therapy with teriparatide and denosumab and denosumab monotherapy in suppressing fragility fracture risk.

In this retrospective study, the effectiveness of short-term teriparatide with denosumab in reducing fragility fracture risk was determined in comparison with denosumab monotherapy. Administration of sequential teriparatide with denosumab showed excellent outcomes in suppressing the risk for fragility fractures compared with denosumab monotherapy. INTRODUCTION: To determine the effectiveness of short-term teriparatide with denosumab in reducing the risk of fragility fractures in comparison to denosumab monotherapy. METHODS: The data of postmenopausal patients treated with denosumab for > 2 years between August 2015 and October 2020 were retrospectively analyzed. One hundred sixty four postmenopausal women of a total 615 were excluded, since they did not undergo > 2 bone mineral density (BMD) tests, were lost to follow-up, or received long-term teriparatide therapy. Total 320 patients received denosumab monotherapy and 131 patients received teriparatide for ≥ 3 months followed by denosumab. The number of osteoporotic fractures, presence of back pain before and after treatment, and annual BMD during treatment were comparatively assessed using t-test, Chi-square test, and linear mixed model analysis. RESULTS: Before treatment, the denosumab monotherapy group had fewer osteoporotic fractures (mean ± standard deviation; 0.459 ± 0.689) than the sequential therapy group had (1.037 ± 0.871; p < 0.001). After treatment, the sequential therapy group had fewer osteoporotic fractures than the denosumab monotherapy group had (0.119 ± 0.348 versus 0.144 ± 0.385; p < 0.001). At 1 and 2 years after treatment, the increase in lumbar spine BMD was greater in the sequential therapy group than in the denosumab monotherapy group (p = 0.08, group × time). The difference between post and pre-treatment back pain visual analog scale score was significantly lower in the sequential therapy group than in the monotherapy group (3.246 ± 3.426 versus 1.734 ± 3.049; p < 0.001). CONCLUSION: Short-term teriparatide use before denosumab showed excellent outcomes in suppressing the risk of fragility fractures compared with denosumab monotherapy.

Programmable microencapsulation for enhanced mesenchymal stem cell persistence and immunomodulation.

Mesenchymal stem cell (MSC) therapies demonstrate particular promise in ameliorating diseases of immune dysregulation but are hampered by short in vivo cell persistence and inconsistencies in phenotype. Here, we demonstrate that biomaterial encapsulation into alginate using a microfluidic device could substantially increase in vivo MSC persistence after intravenous (i.v.) injection. A combination of cell cluster formation and subsequent cross-linking with polylysine led to an increase in injected MSC half-life by more than an order of magnitude. These modifications extended persistence even in the presence of innate and adaptive immunity-mediated clearance. Licensing of encapsulated MSCs with inflammatory cytokine pretransplantation increased expression of immunomodulatory-associated genes, and licensed encapsulates promoted repopulation of recipient blood and bone marrow with allogeneic donor cells after sublethal irradiation by a ∼2-fold increase. The ability of microgel encapsulation to sustain MSC survival and increase overall immunomodulatory capacity may be applicable for improving MSC therapies in general.

Robust Human Activity Recognition by Integrating Image and Accelerometer Sensor Data Using Deep Fusion Network.

Studies on deep-learning-based behavioral pattern recognition have recently received considerable attention. However, if there are insufficient data and the activity to be identified is changed, a robust deep learning model cannot be created. This work contributes a generalized deep learning model that is robust to noise not dependent on input signals by extracting features through a deep learning model for each heterogeneous input signal that can maintain performance while minimizing preprocessing of the input signal. We propose a hybrid deep learning model that takes heterogeneous sensor data, an acceleration sensor, and an image as inputs. For accelerometer data, we use a convolutional neural network (CNN) and convolutional block attention module models (CBAM), and apply bidirectional long short-term memory and a residual neural network. The overall accuracy was 94.8% with a skeleton image and accelerometer data, and 93.1% with a skeleton image, coordinates, and accelerometer data after evaluating nine behaviors using the Berkeley Multimodal Human Action Database (MHAD). Furthermore, the accuracy of the investigation was revealed to be 93.4% with inverted images and 93.2% with white noise added to the accelerometer data. Testing with data that included inversion and noise data indicated that the suggested model was robust, with a performance deterioration of approximately 1%.

Perfusion recovery on TTP maps after endovascular stroke treatment might predict favorable neurological outcomes.

OBJECTIVES: Early recanalization and adequate collateral blood flow are surrogates for functional recovery in endovascular stroke treatment (EVT). We evaluated the prognostic value of pre- and immediate post-thrombectomy perfusion-weighted magnetic resonance imaging (PWI) parameters. METHODS: Consecutive patients with acute ischemic stroke who underwent EVT were enrolled. Lesion volumes and their corresponding changes on diffusion-weighted (DWI) and PWI were assessed. Outcome was measured with modified Rankin Scale (mRS) at 90 days, and early neurological improvement (> 8 points improvement on National Institutes of Health Stroke Scale [NIHSS] or 0 to 1) at 7 days. RESULTS: Fifty-two patients were enrolled. After control of initial NIHSS and recanalization status, post-thrombectomy time-to-peak (TTP) hypoperfused volume and TTP hypoperfused volume change remained independent predictors of favorable functional outcome (odds ratio [OR] = 0.13, 95% confidence interval [CI] = 0.03-0.54, p = 0.005; OR = 1.018, 95% CI = 1.00-1.03, p = 0.017), and early neurological improvement (OR = 0.20, 95% CI 0.07-0.58, p = 0.003; OR = 1.02, 95% CI = 1.00-1.03, p = 0.010). The areas under the curve of post-thrombectomy TTP hypoperfused volume and TTP hypoperfused volume change were 0.90 and 0.82 (cutoff 68 mL and 56 mL) for favorable outcome and 0.86 and 0.82 (cutoff 76 mL and 58 mL) for early neurological improvement, which had better prognostic values than other MR parameters and recanalization grades. CONCLUSIONS: These results suggest a large amount of perfusion recovery on TTP is associated with favorable outcome as well as early neurological improvement after EVT, and may be a useful prognostic marker. KEY POINTS: • A large amount of perfusion recovery on TTP map is associated with favorable outcome and early neurological improvement after EVT. • The best cutoff value for favorable functional outcome was 68 mL for post-EVT TTP hypoperfused volume and 56 mL decrease for TTP hypoperfused volume. • Amount of perfusion recovery on TTP map has better performance on the prediction of favorable functional recovery and early neurological improvement than other diffusion- and perfusion-weighted MRI parameters and recanalization grades.

Roles of microglial membranes in Alzheimer's disease.

The majority of Alzheimer's disease (AD) risk genes are highly and selectively expressed by microglia in the brain. Several of these genes are related to lipid and cholesterol metabolism, lipid synthesis, lipid transport, endocytosis, exocytosis and phagocytosis. Therefore, studying the roles of cellular membrane biophysics in microglial function should improve our understanding of the AD pathology. In this chapter, we discuss how lipid rafts and membrane-cytoskeleton adhesion impact microglial-mediated oxidative stress and clearance of amyloid-ß peptide (Aß). We also discuss potential roles of lipid membrane-bound extracellular vesicles as carriers of pathological factors to promote inflammation and cytotoxicity.

Influenza Screening via Deep Learning Using a Combination of Epidemiological and Patient-Generated Health Data: Development and Validation Study.

BACKGROUND: Screening for influenza in primary care is challenging due to the low sensitivity of rapid antigen tests and the lack of proper screening tests. OBJECTIVE: The aim of this study was to develop a machine learning-based screening tool using patient-generated health data (PGHD) obtained from a mobile health (mHealth) app. METHODS: We trained a deep learning model based on a gated recurrent unit to screen influenza using PGHD, including each patient's fever pattern and drug administration records. We used meteorological data and app-based surveillance of the weekly number of patients with influenza. We defined a single episode as the set of consecutive days, including the day the user was diagnosed with influenza or another disease. Any record a user entered 24 hours after his or her last record was considered to be the start of a new episode. Each episode contained data on the user's age, gender, weight, and at least one body temperature record. The total number of episodes was 6657. Of these, there were 3326 episodes within which influenza was diagnosed. We divided these episodes into 80% training sets (2664/3330) and 20% test sets (666/3330). A 5-fold cross-validation was used on the training set. RESULTS: We achieved reliable performance with an accuracy of 82%, a sensitivity of 84%, and a specificity of 80% in the test set. After the effect of each input variable was evaluated, app-based surveillance was observed to be the most influential variable. The correlation between the duration of input data and performance was not statistically significant (P=.09). CONCLUSIONS: These findings suggest that PGHD from an mHealth app could be a complementary tool for influenza screening. In addition, PGHD, along with traditional clinical data, could be used to improve health conditions.

Carbonyl Compounds Containing Formaldehyde Produced from the Heated Mouthpiece of Tobacco Sticks for Heated Tobacco Products.

Diverse harmful compounds can be emitted during the heating of tobacco sticks for heated tobacco products (HTPs). In this study, the generation of harmful compounds from the filter, instead of tobacco in tobacco sticks, was confirmed. The heat of a heated tobacco product device can be transferred to the tobacco stick filter, resulting in the generation of harmful compounds from the heated filter. Since the heating materials (tobacco consumable) of the tobacco sticks evaluated in this study were different depending on the brand, the harmful compounds emitted from the heated tobacco stick filters were examined by focusing on the carbonyl compounds, using three different tobacco stick parts. Acetaldehyde and propionaldehyde exhibited the highest concentrations in HTP aerosols produced by heating the tobacco consumable (conventional case) (63.5 ± 18.4 µg/stick and 1.71 ± 0.123 µg/stick, respectively). The aerosols produced by heating tobacco stick filters had higher formaldehyde and acrolein concentrations (0.945 ± 0.214 µg/stick and 0.519 ± 0.379 µg/stick) than the aerosols generated from heated tobacco consumable (0.641 ± 0.092 µg/stick and 0.220 ± 0.102 µg/stick). As such, formaldehyde and acrolein were produced by heating small parts of the mouthpiece of a tobacco stick, regardless of the heated tobacco product brand. In addition, acetone was only detected in the aerosols generated from heated filters (0.580 ± 0.305 µg/stick). Thus, safety evaluations of heated tobacco products should include considerations of the harmful compounds generated by heating tobacco stick mouthpieces for heated tobacco products in addition to those found in heated tobacco product aerosols.

Extracellular matrix stiffness causes systematic variations in proliferation and chemosensitivity in myeloid leukemias.

Extracellular matrix stiffness influences biological functions of some tumors. However, it remains unclear how cancer subtypes with different oncogenic mutations respond to matrix stiffness. In addition, the relevance of matrix stiffness to in vivo tumor growth kinetics and drug efficacy remains elusive. Here, we designed 3D hydrogels with physical parameters relevant to hematopoietic tissues and adapted them to a quantitative high-throughput screening format to facilitate mechanistic investigations into the role of matrix stiffness on myeloid leukemias. Matrix stiffness regulates proliferation of some acute myeloid leukemia types, including MLL-AF9+ MOLM-14 cells, in a biphasic manner by autocrine regulation, whereas it decreases that of chronic myeloid leukemia BCR-ABL+ K-562 cells. Although Arg-Gly-Asp (RGD) integrin ligand and matrix softening confer resistance to a number of drugs, cells become sensitive to drugs against protein kinase B (PKB or AKT) and rapidly accelerated fibrosarcoma (RAF) proteins regardless of matrix stiffness when MLL-AF9 and BCR-ABL are overexpressed in K-562 and MOLM-14 cells, respectively. By adapting the same hydrogels to a xenograft model of extramedullary leukemias, we confirm the pathological relevance of matrix stiffness in growth kinetics and drug sensitivity against standard chemotherapy in vivo. The results thus demonstrate the importance of incorporating 3D mechanical cues into screening for anticancer drugs.

Deterministic encapsulation of single cells in thin tunable microgels for niche modelling and therapeutic delivery.

Existing techniques to encapsulate cells into microscale hydrogels generally yield high polymer-to-cell ratios and lack control over the hydrogel's mechanical properties. Here, we report a microfluidic-based method for encapsulating single cells in an approximately six-micrometre layer of alginate that increases the proportion of cell-containing microgels by a factor of ten, with encapsulation efficiencies over 90%. We show that in vitro cell viability was maintained over a three-day period, that the microgels are mechanically tractable, and that, for microscale cell assemblages of encapsulated marrow stromal cells cultured in microwells, osteogenic differentiation of encapsulated cells depends on gel stiffness and cell density. We also show that intravenous injection of singly encapsulated marrow stromal cells into mice delays clearance kinetics and sustains donor-derived soluble factors in vivo. The encapsulation of single cells in tunable hydrogels should find use in a variety of tissue engineering and regenerative medicine applications.

Are glass fiber particles released during the use of electronic cigarettes? Development of a semi-quantitative approach to detect glass particle emission due to vaping.

This study investigated the emission characteristics of glass particles resulting from smoking electronic cigarettes (ECs). First, the most suitable filter for the collection of glass particles was explored by examining the performance (reliability) of various types of filters. A polycarbonate filter was determined as the optimum choice to collect glass particles in EC aerosol. A cartomizer was filled with EC refill solution composed of an equal volume of propylene glycol (PG) and vegetable glycol (VG). To simulate the potential conditions for glass particle emission, EC vaped aerosols were collected at three distinctive puffing intervals: (1) 0-10 puffs, (2) 101-110 puffs, and (3) 201-210 puffs (flow rate of 1 L min-1, 2 s per puff, and 10 puffs per sample). Glass particles were observed as early as after 100 times puffing from certain products, while after 200 from others. Thus, glass particles were generated by increasing the number of puffs and usage of the EC cartomizer. The analysis of glass particles collected onto polycarbonate filters by scanning electron microscopy (SEM)/energy dispersive X-ray spectroscopy (EDS) confirmed the presence of glass particles in samples collected after puffing 100-200 times. The study demonstrated that the possibility of glass particle emissions from the EC device increased considerably with the increasing number of total puffs.

Myosin-II repression favors pre/proplatelets but shear activation generates platelets and fails in macrothrombocytopenia.

Megakaryocyte ploidy and the generation of pre/proplatelets are both increased in culture by pharmacologic inhibition of myosin-II, but nonmuscle myosin-IIA (MIIA) mutations paradoxically cause MYH9-related diseases (MYH9-RD) that adversely affect platelets. In marrow, megakaryocytes extend projections into the microcirculation, where shear facilitates fragmentation to large pre/proplatelets, suggesting that fluid stresses and myosin-II activity somehow couple in platelet biogenesis. Here, in bulk shear, plateletlike particles generated from megakaryocytes are maximized at a shear stress typical of that in the microcirculation and after treatment with a myosin-II inhibitor. MIIA activity in static cells is naturally repressed through phosphorylation at Serine-1943, but shear decreases phosphorylation, consistent with MIIA activation and localization to platelet cortex. Micropipette aspiration of cells shows myosin-II accumulates at stressed sites, but its inhibition prevents such mechanoactivation and facilitates generation of CD41(+) fragments similar in size to pre/proplatelets. MYH9-RD mutants phenocopy inhibition, revealing a dominant negative effect. MIIA is diffuse in the large platelets of a MYH9-RD patient with macrothrombocytopenia and is also diffuse in normal pre/proplatelets treated with inhibitor that blocks in vitro division to small platelets. The findings explain the large platelets in MYH9-RD and the near-normal thrombocrit of patients. Myosin-II regulation thus controls platelet size and number.

Blood and immune cell engineering: Cytoskeletal contractility and nuclear rheology impact cell lineage and localization: Biophysical regulation of hematopoietic differentiation and trafficking.

Clinical success with human hematopoietic stem cell (HSC) transplantation establishes a paradigm for regenerative therapies with other types of stem cells. However, it remains generally challenging to therapeutically treat tissues after engineering of stem cells in vitro. Recent studies suggest that stem and progenitor cells sense physical features of their niches. Here, we review biophysical contributions to lineage decisions, maturation, and trafficking of blood and immune cells. Polarized cellular contractility and nuclear rheology are separately shown to be functional markers of a hematopoietic hierarchy that predict the ability of a lineage to traffic in and out of the bone marrow niche. These biophysical determinants are regulated by a set of structural molecules, including cytoplasmic myosin-II and nuclear lamins, which themselves are modulated by a diverse range of transcriptional and post-translational mechanisms. Small molecules that target these mechanobiological circuits, along with novel bioengineering methods, could prove broadly useful in programming blood and immune cells for therapies ranging from blood transfusions to immune attack of tumors.

Lamins regulate cell trafficking and lineage maturation of adult human hematopoietic cells.

Hematopoietic stem and progenitor cells, as well as nucleated erythroblasts and megakaryocytes, reside preferentially in adult marrow microenvironments whereas other blood cells readily cross the endothelial barrier into the circulation. Because the nucleus is the largest organelle in blood cells, we hypothesized that (i) cell sorting across microporous barriers is regulated by nuclear deformability as controlled by lamin-A and -B, and (ii) lamin levels directly modulate hematopoietic programs. Mass spectrometry-calibrated intracellular flow cytometry indeed reveals a lamin expression map that partitions human blood lineages between marrow and circulating compartments (P = 0.00006). B-type lamins are highly variable and predominate only in CD34(+) cells, but migration through micropores and nuclear flexibility in micropipette aspiration both appear limited by lamin-A:B stoichiometry across hematopoietic lineages. Differentiation is also modulated by overexpression or knockdown of lamins as well as retinoic acid addition, which regulates lamin-A transcription. In particular, erythroid differentiation is promoted by high lamin-A and low lamin-B1 expression whereas megakaryocytes of high ploidy are inhibited by lamin suppression. Lamins thus contribute to both trafficking and differentiation.