Programmable 3D silk bone marrow niche for platelet generation ex vivo and modeling of megakaryopoiesis pathologies.
AuthorsDi Buduo CA, Wray LS, Tozzi L, Malara A, Chen Y, Ghezzi CE, Smoot D, Sfara C, Antonelli A, Spedden E, Bruni G, Staii C, De Marco L, Magnani M, Kaplan DL, Balduini A,
Journal
PubMed ID25575540
'We present a programmable bioengineered three-dimensional silk based bone marrow niche tissue system that successfully mimics the physiology of human bone marrow environment allowing us to manufacture functional human platelets ex vivo. Using stem/progenitor cells, megakaryocyte function and platelet generation were recorded in response to variations in extracellular matrix components, ... More
Regulation of Phagocytosis in Macrophages by Membrane Ethanolamine Plasmalogens.
'Macrophages, as professional phagocytes of the immune system, possess the ability to detect and clear invading pathogens and apoptotic cells through phagocytosis. Phagocytosis involves membrane reorganization and remodeling events on the cell surface, which play an essential role in innate immunity and tissue homeostasis and the control of inflammation. In ... More
Nano-scale microfluidics to study 3D chemotaxis at the single cell level.
AuthorsFrick C, Dettinger P, Renkawitz J, Jauch A, Berger CT, Recher M, Schroeder T, Mehling M,
JournalPLoS One
PubMed ID29879160
'Directed migration of cells relies on their ability to sense directional guidance cues and to interact with pericellular structures in order to transduce contractile cytoskeletal- into mechanical forces. These biomechanical processes depend highly on microenvironmental factors such as exposure to 2D surfaces or 3D matrices. In vivo, the majority of ... More
Tumor-stroma interactions differentially alter drug sensitivity based on the origin of stromal cells.
'Due to tumor heterogeneity, most believe that effective treatments should be tailored to the features of an individual tumor or tumor subclass. It is still unclear, however, what information should be considered for optimal disease stratification, and most prior work focuses on tumor genomics. Here, we focus on the tumor ... More
Neutrophil-generated HOCl leads to non-specific thiol oxidation in phagocytized bacteria.
AuthorsDegrossoli A, Müller A, Xie K, Schneider JF, Bader V, Winklhofer KF, Meyer AJ, Leichert LI,
JournalElife
PubMed ID29506649
'Phagocytic immune cells kill pathogens in the phagolysosomal compartment with a cocktail of antimicrobial agents. Chief among them are reactive species produced in the so-called oxidative burst. Here, we show that bacteria exposed to a neutrophil-like cell line experience a rapid and massive oxidation of cytosolic thiols. Using roGFP2-based fusion ... More
'The purpose of this study was to characterize whether induced pluripotent stem cells (iPSCs) affect survival of grafted retinal ganglion cells (RGCs) after transplantation. For in vitro studies, human iPSCs were either directly cocultured with mouse RGCs or plated in hanging inserts in RGC cultures for 1 week. For ex ... More
Sensing of HIV-1 by TLR8 activates human T cells and reverses latency.
AuthorsMeås HZ, Haug M, Beckwith MS, Louet C, Ryan L, Hu Z, Landskron J, Nordbø SA, Taskén K, Yin H, Damås JK, Flo TH
JournalNat Commun
PubMed ID31919342
'During HIV infection, cell-to-cell transmission results in endosomal uptake of the virus by target CD4+ T cells and potential exposure of the viral ssRNA genome to endosomal Toll-like receptors (TLRs). TLRs are instrumental in activating inflammatory responses in innate immune cells, but their function in adaptive immune cells is less well ... More
Detection and prognostic role of heterogeneous populations of melanoma circulating tumour cells.
AuthorsAya-Bonilla CA, Morici M, Hong X, McEvoy AC, Sullivan RJ, Freeman J, Calapre L, Khattak MA, Meniawy T, Millward M, Ziman M, Gray ES
JournalBr J Cancer
PubMed ID32037400
'Circulating tumour cells (CTCs) can be assessed through a minimally invasive blood sample with potential utility as a predictive, prognostic and pharmacodynamic biomarker. The large heterogeneity of melanoma CTCs has hindered their detection and clinical application.' ... More
The AMP-activated protein kinase beta 1 subunit modulates erythrocyte integrity.
AuthorsCambridge EL, McIntyre Z, Clare S, Arends MJ, Goulding D, Isherwood C, Caetano SS, Reviriego CB, Swiatkowska A, Kane L, Harcourt K, Adams DJ, White JK, Speak AO,
JournalExp Hematol
PubMed ID27666489
'Failure to maintain a normal in vivo erythrocyte half-life results in the development of hemolytic anemia. Half-life is affected by numerous factors, including energy balance, electrolyte gradients, reactive oxygen species, and membrane plasticity. The heterotrimeric AMP-activated protein kinase (AMPK) is an evolutionarily conserved serine/threonine kinase that acts as a critical regulator ... More
Engineered proteins with sensing and activating modules for automated reprogramming of cellular functions.
AuthorsSun J, Lei L, Tsai CM, Wang Y, Shi Y, Ouyang M, Lu S, Seong J, Kim TJ, Wang P, Huang M, Xu X, Nizet V, Chien S, Wang Y,
JournalNat Commun
PubMed ID28883531
'Protein-based biosensors or activators have been engineered to visualize molecular signals or manipulate cellular functions. Here we integrate these two functionalities into one protein molecule, an integrated sensing and activating protein (iSNAP). A prototype that can detect tyrosine phosphorylation and immediately activate auto-inhibited Shp2 phosphatase, Shp2-iSNAP, is designed through modular ... More
Combined toll-like receptor 3/7/9 deficiency on host cells results in T-cell-dependent control of tumour growth.
AuthorsKlein JC, Moses K, Zelinskyy G, Sody S, Buer J, Lang S, Helfrich I, Dittmer U, Kirschning CJ, Brandau S,
JournalNat Commun
PubMed ID28300057
'Toll-like receptors (TLRs) are located either on the cell surface or intracellularly in endosomes and their activation normally contributes to the induction of protective immune responses. However, in cancer their activation by endogenous ligands can modulate tumour progression. It is currently unknown how endosomal TLRs regulate endogenous anti-tumour immunity. Here ... More
Innate immune activating ligand SUMOylation affects tumor cell recognition by NK cells.
AuthorsZitti B, Molfetta R, Fionda C, Quatrini L, Stabile H, Lecce M, de Turris V, Ricciardi MR, Petrucci MT, Cippitelli M, Gismondi A, Santoni A, Paolini R,
JournalSci Rep
PubMed ID28874810
'Natural Killer cells are innate lymphocytes involved in tumor immunosurveillance. They express activating receptors able to recognize self-molecules poorly expressed on healthy cells but up-regulated upon stress conditions, including transformation. Regulation of ligand expression in tumor cells mainly relays on transcriptional mechanisms, while the involvement of ubiquitin or ubiquitin-like modifiers ... More
Combining mechanical foaming and thermally induced phase separation to generate chitosan scaffolds for soft tissue engineering.
AuthorsBiswas DP, Tran PA, Tallon C, O'Connor AJ,
JournalJ Biomater Sci Polym Ed
PubMed ID27875928
'In this paper, a novel foaming methodology consisting of turbulent mixing and thermally induced phase separation (TIPS) was used to generate scaffolds for tissue engineering. Air bubbles were mechanically introduced into a chitosan solution which forms the continuous polymer/liquid phase in the foam created. The air bubbles entrained in the ... More
The microprotein Minion controls cell fusion and muscle formation.
AuthorsZhang Q, Vashisht AA, O'Rourke J, Corbel SY, Moran R, Romero A, Miraglia L, Zhang J, Durrant E, Schmedt C, Sampath SC, Sampath SC,
JournalNat Commun
PubMed ID28569745
'Although recent evidence has pointed to the existence of small open reading frame (smORF)-encoded microproteins in mammals, their function remains to be determined. Skeletal muscle development requires fusion of mononuclear progenitors to form multinucleated myotubes, a critical but poorly understood process. Here we report the identification of Minion (microprotein inducer ... More
Intravital Microscopy Analysis of Hepatic T Cell Dynamics.
AuthorsBenechet AP, Ganzer L, Iannacone M,
JournalMethods Mol Biol
PubMed ID27787791
T cells play critical roles in controlling hepatotropic viral infections and liver tumors. The protective capacity of these cells is mediated by antigen-experienced effector cells and depends on their ability to migrate to and traffic within the liver, recognize pathogen- or tumor-derived antigens, get activated and deploy effector functions.While some ... More
Automated microraft platform to identify and collect non-adherent cells successfully gene-edited with CRISPR-Cas9.
Microraft arrays have been used to screen and then isolate adherent and non-adherent cells with very high efficiency and excellent viability; however, manual screening and isolation limits the throughput and utility of the technology. In this work, novel hardware and software were developed to automate the microraft array platform. The ... More
Presence of diabetes autoantigens in extracellular vesicles derived from human islets.
AuthorsHasilo CP, Negi S, Allaeys I, Cloutier N, Rutman AK, Gasparrini M, Bonneil É, Thibault P, Boilard É, Paraskevas S,
JournalSci Rep
PubMed ID28694505
Beta-cell (ß-cell) injury is the hallmark of autoimmune diabetes. However, the mechanisms by which autoreactive responses are generated in susceptible individuals are not well understood. Extracellular vesicles (EV) are produced by mammalian cells under normal and stressed physiological states. They are an important part of cellular communication, and may serve ... More
Disruption of outer blood-retinal barrier by Toxoplasma gondii-infected monocytes is mediated by paracrinely activated FAK signaling.
AuthorsSong HB, Jun HO, Kim JH, Lee YH, Choi MH, Kim JH,
JournalPLoS One
PubMed ID28406972
Ocular toxoplasmosis is mediated by monocytes infected with Toxoplasma gondii that are disseminated to target organs. Although infected monocytes can easily access to outer blood-retinal barrier due to leaky choroidal vasculatures, not much is known about the effect of T. gondii-infected monocytes on outer blood-retinal barrier. We prepared human monocytes, ... More
Sensitive and predictable separation of microfluidic droplets by size using in-line passive filter.
AuthorsDing R, Ung WL, Heyman JA, Weitz DA,
JournalBiomicrofluidics
PubMed ID28344725
Active manipulation of droplets is crucial in droplet microfluidics. However, droplet polydispersity decreases the accuracy of active manipulation. We develop a microfluidic
Printed droplet microfluidics for on demand dispensing of picoliter droplets and cells.
Although the elementary unit of biology is the cell, high-throughput methods for the microscale manipulation of cells and reagents are limited. The existing options either are slow, lack single-cell specificity, or use fluid volumes out of scale with those of cells. Here we present printed droplet microfluidics, a technology to ... More
A subset of extracellular vesicles carries the bulk of microRNAs in commercial dairy cow's milk.
AuthorsBenmoussa A, Ly S, Shan ST, Laugier J, Boilard E, Gilbert C, Provost P,
JournalJ Extracell Vesicles
PubMed ID29904572
MicroRNAs are small gene-regulatory RNAs that are found in various biological fluids, including milk, where they are often contained inside extracellular vesicles (EVs), like exosomes. In a previous study, we reported that commercial dairy cow's milk microRNAs resisted simulated digestion and were not exclusively associated with canonical exosomes. Here, we ... More
Label-Free Detection of Microvesicles and Proteins by the Bundling of Gliding Microtubules.
AuthorsChaudhuri S, Korten T, Korten S, Milani G, Lana T, Te Kronnie G, Diez S,
JournalNano Lett
PubMed ID29202578
Development of miniaturized devices for the rapid and sensitive detection of analyte is crucial for various applications across healthcare, pharmaceutical, environmental, and other industries. Here, we report on the detection of unlabeled analyte by using fluorescently labeled, antibody-conjugated microtubules in a kinesin-1 gliding motility assay. The detection principle is based ... More
Recombinant phosphatidylserine-binding nanobodies for targeting of extracellular vesicles to tumor cells: a plug-and-play approach.
AuthorsKooijmans SAA, Gitz-Francois JJJM, Schiffelers RM, Vader P,
JournalNanoscale
PubMed ID29334397
Extracellular vesicles (EVs) are increasingly being recognized as candidate drug delivery systems due to their ability to functionally transfer biological cargo between cells. However, manipulation of targeting properties of EVs through engineering of the producer cells can be challenging and time-consuming. As a novel approach to confer tumor targeting properties ... More
Coactivation of TLR2 and TLR8 in Primary Human Monocytes Triggers a Distinct Inflammatory Signaling Response.
AuthorsBösl K, Giambelluca M, Haug M, Bugge M, Espevik T, Kandasamy RK, Bergstrøm B,
JournalFront Physiol
PubMed ID29896111
Innate immune signaling is essential to mount a fast and specific immune response to pathogens. Monocytes and macrophages are essential cells in the early response in their capacity as ubiquitous phagocytic cells. They phagocytose microorganisms or damaged cells and sense pathogen/damage-associated molecular patterns (PAMPs/DAMPs) through innate receptors such as Toll-like ... More
CD5L Promotes M2 Macrophage Polarization through Autophagy-Mediated Upregulation of ID3.
AuthorsSanjurjo L, Aran G, Téllez É, Amézaga N, Armengol C, López D, Prats C, Sarrias MR,
JournalFront Immunol
PubMed ID29593730
CD5L (CD5 molecule-like) is a secreted glycoprotein that controls key mechanisms in inflammatory responses, with involvement in processes such as infection, atherosclerosis, and cancer. In macrophages, CD5L promotes an anti-inflammatory cytokine profile in response to TLR activation. In the present study, we questioned whether CD5L is able to influence human ... More
Transient non-integrative expression of nuclear reprogramming factors promotes multifaceted amelioration of aging in human cells.
AuthorsSarkar TJ, Quarta M, Mukherjee S, Colville A, Paine P, Doan L, Tran CM, Chu CR, Horvath S, Qi LS, Bhutani N, Rando TA, Sebastiano V
JournalNat Commun
PubMed ID32210226
Aging is characterized by a gradual loss of function occurring at the molecular, cellular, tissue and organismal levels. At the chromatin level, aging associates with progressive accumulation of epigenetic errors that eventually lead to aberrant gene regulation, stem cell exhaustion, senescence, and deregulated cell/tissue homeostasis. Nuclear reprogramming to pluripotency can ... More
Co-culture of induced pluripotent stem cells with cardiomyocytes is sufficient to promote their differentiation into cardiomyocytes.
AuthorsChu AJ, Zhao EJ, Chiao M, Lim CJ
JournalPLoS One
PubMed ID32243463
Various types of stem cells and non-stem cells have been shown to differentiate or transdifferentiate into cardiomyocytes by way of co-culture with appropriate inducer cells. However, there is a limited demonstration of a co-culture induction system utilizing stem cell-derived cardiomyocytes as a stimulatory source for cardiac reprogramming (of stem cells ... More
A Platform for Studying of the Three-Dimensional Migration of Hematopoietic Stem/Progenitor Cells.
AuthorsLee E, Kim J, Kang Y, Shin JW
JournalTissue Eng Regen Med
PubMed ID32002840
Hematopoietic stem/progenitor cells (HSPCs) have the property to return to the bone marrow, which is believed to be critical in situations such as HSPC transplantation. This property plays an important role in the stemness, viability, and proliferation of HSPCs, also. However, most in vitro models so far have not sufficiently ... More
C3a elicits unique migratory responses in immature low-density neutrophils.
AuthorsHsu BE, Roy J, Mouhanna J, Rayes RF, Ramsay L, Tabariès S, Annis MG, Watson IR, Spicer JD, Costantino S, Siegel PM
JournalOncogene
PubMed ID32020055
Neutrophils represent the immune system's first line of defense and are rapidly recruited into inflamed tissue. In cancer associated inflammation, phenotypic heterogeneity has been ascribed to this cell type, whereby neutrophils can manifest anti- or pro-metastatic functions depending on the cellular/micro-environmental context. Here, we demonstrate that pro-metastatic immature low-density neutrophils ... More
Tumor-treating fields (TTFields) induce immunogenic cell death resulting in enhanced antitumor efficacy when combined with anti-PD-1 therapy.
AuthorsVoloshin T, Kaynan N, Davidi S, Porat Y, Shteingauz A, Schneiderman RS, Zeevi E, Munster M, Blat R, Tempel Brami C, Cahal S, Itzhaki A, Giladi M, Kirson ED, Weinberg U, Kinzel A, Palti Y
JournalCancer Immunol Immunother
PubMed ID32144446
Tumor-treating fields (TTFields) are alternating electric fields in a specific frequency range (100-300 kHz) delivered to the human body through transducer arrays. In this study, we evaluated whether TTFields-mediated cell death can elicit antitumoral immunity and hence would be effectively combined with anti-PD-1 therapy. We demonstrate that in TTFields-treated cancer cells, ... More
Inhibition of miR30a-3p by sulforaphane enhances gap junction intercellular communication in pancreatic cancer.
AuthorsGeorgikou C, Yin L, Gladkich J, Xiao X, Sticht C, Torre C, Gretz N, Gross W, Schäfer M, Karakhanova S, Herr I
JournalCancer Lett
PubMed ID31678166
The therapy resistance of pancreatic cancer is associated with the loss of gap junction intercellular communication and connexin 43 expression. The broccoli isothiocyanate sulforaphane restored these features and therapy sensitivity. We investigated whether microRNA signaling is involved. Established cell lines and a patient tissue array (n?=?96) were evaluated by miRNA ... More