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Biology subjects

Walz, K.

Publications and source records attributed to Walz, K..

5 recordsLinked to original sources

Human knee osteoarthritis patient-specific cartilage-on-a-chip model captures donor differences to stressors and treatments

Knee Osteoarthritis (KOA) is a progressive whole-joint disease without approved disease modifying OA drugs (DMOADs). Effective treatments have been hindered by multiple layers of heterogeneity, including diverse disease etiology and patient-to-patient variability. Here, we present a scalable, KOA patient-derived (PD) cartilage-on-a-chip (CartChip) model integrating end-stage KOA cartilage tissue explants on a microengineered platform that, under mechanical overloading and hyperinflammatory stressors, mimics different KOA etiologies. These stressors drove distinct multivariable model features, including changes in a curated panel of anabolic and catabolic genes, extracellular matrix protein and soluble factors. Exploratory analysis of coordinated model readouts identified stressor-agnostic and -specific KOA disease signatures. Despite using KOA tissue, the model showed improvements to dexamethasone, a symptom-modifying, anti-inflammatory KOA treatment. The model responses to dexamethasone were dependent on both stressor and donor heterogeneity. Exploratory groupings of coordinated model readouts provided proof-of-concept for predicting categories of patient responsiveness to test therapeutics. Annotating patient data provided additional donor-dependent contexts for interpreting model responsiveness. PD-CartChip provides a powerful research platform to potentially surmount the donor and stressor heterogeneity barrier in developing DMOADs.

bioengineering↗

Connecting adhesion dynamics and trail formation in malaria parasites by imaging the major surface antigens CSP and TRAP

Malaria infections are initiated by mosquito bites, during which Plasmodium sporozoites are injected into the host skin. Sporozoites migrate rapidly to find and enter blood capillaries and ultimately invade hepatocytes. Sporozoite migration and invasion is mediated by the transmembrane protein thrombospondin-related anonymous protein (TRAP), which links the extracellular substrate to the actomyosin complex powering gliding motility, while the abundant, GPI-anchored circumsporozoite protein (CSP) covers most of the parasite membrane and modulates adhesion. Both proteins are secreted onto the parasite surface and deposited in a membranous trail originating at the parasite rear. The surface dynamics of these essential sporozoite proteins and the mechanism of deposition, however, are not understood. Here, using orbital total internal reflection fluorescence microscopy (TIRF), we reveal the dynamics of TRAP adhesion site formation and disassembly as well as CSP and TRAP deposition rates. We find that TRAP assembles into distinct adhesion sites, which then undergo retrograde translocation as the sporozoite moves forward. Around half of the TRAP adhesins, together with CSP, remain associated in small membrane droplets on the substrate after the sporozoite has disengaged from the adhesion site. These droplets seem to originate from nanotubes, that presumably decay under high tension. Strikingly, we observe a change in actin filament accumulation if proteolytic cleavage of TRAP is inhibited, providing the first visual evidence for outside-in signaling in sporozoites. Our study reveals a relation between adhesion dynamics and trail formation in Plasmodium sporozoites that might also be relevant for other cell types.

cell biology↗

Dysregulation of cell migration by matrix metalloproteinases in geleophysic dysplasia

Geleophysic dysplasia (GD) is characterized by short stature, brachydactyly, joint limitations, a distinctive facial appearance, as well as cardiac and respiratory dysfunction that can be life-threatening. GD is caused by pathogenic variants in the ADAMTSL2, FBN1, or LTBP3 genes. While dermal fibroblasts derived from affected individuals have shown poor organization of the extracellular matrix (ECM), it remains elusive how the disorganized ECM contributes to GD pathogenesis. To understand the molecular mechanisms in GD, we isolated and characterized primary human dermal fibroblasts from affected individuals with ADAMTSL2 and FBN1 variants. We found that the secretion of ECM proteins including ADAMTSL2, FBN1, and Fibronectin were impaired in GD fibroblasts. Increased cell migration was observed in GD fibroblasts carrying ADAMTSL2 or FBN1 variants, which was associated with up-regulation of MMP-1 and MMP-14, two proteases related to cell mobility. The enhanced cell migration and up-regulation of MMP-1 and MMP-14 were corroborated in mouse primary dermal fibroblasts carrying pathogenic variants in Adamtsl2 and in lung and heart tissues from Adamtsl2-knockout mice. A pan MMP inhibitor, GM6001, inhibited the migration of GD fibroblasts. Overall, our results suggest that MMP-1/-14 up-regulation play a role in the development of GD and may be utilized as a treatment target.

cell biology↗

B cells targeting parasites capture spatially linked antigens to secure T cell help

Our understanding of T-cell-dependent humoral responses has been largely shaped by studies involving model antigens such as recombinant proteins and viruses 1,2. In these contexts, B cells internalize the entire antigen or pathogen, and present a range of antigens to helper CD4+ T cells to initiate the humoral response. However, this model does not account for large pathogens (such as parasites) that are too large to be taken up by individual B cells, and the mechanisms by which B cells acquire and present antigens from large complex pathogens to T cells remain poorly understood. Here we used Plasmodium, the causative parasite of malaria, as a model to investigate the requirements for T cell help for B cells targeting the Plasmodium surface circumsporozoite protein (CSP). Upon Plasmodium sporozoite (SPZ) immunization, CSP-specific B cells can form a synapse-like structure with SPZs and take up CSP and non-CSP surface antigens. As a result, CSP-specific B cells can receive help from CD4+ T cells specific to antigens that are located on the surface but not cytosol of the Plasmodium SPZ. Therefore, B cells can obtain help, not only from T cells with the same protein specificity, but also from T cells specific for spatially linked antigens. This flexibility in T cell help may enhance the initiation and maintenance of humoral immune responses to complex pathogens.

immunology↗

Sord deficient rats develop a motor-predominant peripheral neuropathy unveiling novel pathophysiological insights

Biallelic SORD mutations cause one of the most frequent forms of recessive hereditary neuropathy, estimated to affect approximately 10,000 patients in North America and Europe alone. Pathogenic SORD loss-of-function changes in the encoded enzyme sorbitol dehydrogenase result in abnormally high sorbitol levels in cells and serum. How sorbitol accumulation leads to peripheral neuropathy remains to be elucidated. A reproducible animal model for SORD neuropathy is essential to illuminate the pathogenesis of SORD deficiency and for preclinical studies of potential therapies. Therefore, we have generated a Sord knockout (KO), Sord-/-, Sprague Dawley rat, to model the human disease and to investigate the pathophysiology underlying SORD deficiency. We have characterized the phenotype in these rats with a battery of behavioral tests as well as biochemical, physiological, and comprehensive histological examinations. Sord-/- rats had remarkably increased levels of sorbitol in serum, cerebral spinal fluid (CSF), and peripheral nerve. Moreover, serum from Sord-/- rats contained significantly increased levels of neurofilament light chain, NfL, an established biomarker for axonal degeneration. Motor performance significantly declined in Sord-/- animals starting at [~]7 months of age. Gait analysis evaluated with video motion tracking confirmed abnormal gait patterns in the hindlimbs. Motor nerve conduction velocities of the tibial nerves were slowed. Light and electron microscopy of the peripheral nervous system revealed degenerating myelinated axons, de- and remyelinated axons, and a likely pathognomonic finding - enlarged "ballooned" myelin sheaths. These findings mainly affected myelinated motor axons; myelinated sensory axons were largely spared. In summary, Sord-/- rats develop a motor-predominant neuropathy that closely resembles the human phenotype. Our studies revealed novel significant aspects of SORD deficiency, and this model will lead to an improved understanding of the pathophysiology and the therapeutic options for SORD neuropathy.

neuroscience↗