ADAMTS Proteins and Vascular Remodeling in Aortic Aneurysms

Extracellular matrix (ECM) in the vascular wall is a highly dynamic structure composed of a set of different molecules such as elastins, collagens, fibronectin (Fn), laminins, proteoglycans, and polysaccharides. ECM undergoes remodeling processes to regulate vascular smooth muscle and endothelial cells’ proliferation, differentiation, and adhesion. Abnormalities affecting the ECM can lead to alteration in cellular behavior and from this, this can conduce to the development of pathologies. Metalloproteases play a key role in maintaining the homeostasis of ECM by mediating the cleavage of different ECM components. There are different types of metalloproteases: matrix metalloproteinases (MMPs), disintegrin and metalloproteinases (ADAMs), and ADAMs with thrombospondin motifs (ADAMTSs). ADAMTSs have been found to participate in cardiovascular physiology and diseases and specifically in aortic aneurysms. This review aims to decipher the potential role of ADAMTS proteins in the physiopathologic development of Thoracic Aortic Aneurysms (TAA) and Abdominal Aortic Aneurysms (AAA). This review will focus on what is known on the ADAMTS family involved in human aneurysms from human tissues to mouse models. The recent findings on THSD4 (encoding ADAMTSL6) mutations in TAA give a new insight on the involvement of the ADAMTS family in TAA.

Mapping information-rich genotype-phenotype landscapes with genome-scale Perturb-seq

A central goal of genetics is to define the relationships between genotypes and phenotypes. High-content phenotypic screens such as Perturb-seq (pooled CRISPR-based screens with single-cell RNA-sequencing readouts) enable massively parallel functional genomic mapping but, to date, have been used at limited scales. Here, we perform genome-scale Perturb-seq targeting all expressed genes with CRISPR interference (CRISPRi) across >2.5 million human cells and present a framework to power biological discovery with the resulting genotype-phenotype map. We use transcriptional phenotypes to predict the function of poorly-characterized genes, uncovering new regulators of ribosome biogenesis (including CCDC86, ZNF236, and SPATA5L1), transcription (C7orf26), and mitochondrial respiration (TMEM242). In addition to assigning gene function, single-cell transcriptional phenotypes allow for in-depth dissection of complex cellular phenomena - from RNA processing to differentiation. We leverage this ability to systematically identify the genetic drivers and consequences of aneuploidy and to discover an unanticipated layer of stress-specific regulation of the mitochondrial genome. Our information-rich genotype-phenotype map reveals a multidimensional portrait of gene function and cellular behavior.

Learning Single-Cell Perturbation Responses using Neural Optimal Transport

Understanding and predicting molecular responses towards external perturbations is a core question in molecular biology. Technological advancements in the recent past have enabled the generation of high-resolution single-cell data, making it possible to profile individual cells under different experimentally controlled perturbations. However, cells are typically destroyed during measurement, resulting in unpaired distributions over either perturbed or non-perturbed cells. Leveraging the theory of optimal transport and the recent advents of convex neural architectures, we learn a coupling describing the response of cell populations upon perturbation, enabling us to predict state trajectories on a single-cell level. We apply our approach, CellOT, to predict treatment responses of 21,650 cells subject to four different drug perturbations. CellOT outperforms current state-of-the-art methods both qualitatively and quantitatively, accurately capturing cellular behavior shifts across all different drugs.

A Toolkit for Manipulating Cellular Behavior: Peptide with Tryptophan-Selective Ru-TAP Complex Regioselectively Photolabeling Specific Protein in Live Cell

Using a chemical approach to crosslink functionally versatile bioeffectors (such as peptides) to native proteins of interest (POI) directly inside a living cell is a useful toolbox for chemical biologists. However, this goal has not been reached due to unsatisfactory chemoselectivity, regioselectivity, and protein-selectivity in in-cellulo protein labeling. Herein we report a highly selective photoaffinity labeling (PAL) method using a tryptophan-specific Ru-TAP complex as photocrosslinker (Trp-tag). Aside from the high selectivity, the PAL is blue light driven by a photoinduced electron transfer (PeT) and allows the bioeffector to bear an additional UV-responsive unit. The two different photosensitivities are demonstrated by blue light photocrosslinking a UV-sensitive peptide to POI. The remote-control functionality of the peptide allows POI inhibition after blue light irradiation, and reactivation upon UV photolysis. Cytoskeletal dynamics regulation is demonstrated via the unprecedented in-cellulo POI photomanipulation, which opens a new avenue to endogenous protein modification for novel functions.

Viscoelasticity Acts as a Marker for Tumor Extracellular Matrix Characteristics

Biological materials such as extracellular matrix scaffolds, cancer cells, and tissues are often assumed to respond elastically for simplicity; the viscoelastic response is quite commonly ignored. Extracellular matrix mechanics including the viscoelasticity has turned out to be a key feature of cellular behavior and the entire shape and function of healthy and diseased tissues, such as cancer. The interference of cells with their local microenvironment and the interaction among different cell types relies both on the mechanical phenotype of each involved element. However, there is still not yet clearly understood how viscoelasticity alters the functional phenotype of the tumor extracellular matrix environment. Especially the biophysical technologies are still under ongoing improvement and further development. In addition, the effect of matrix mechanics in the progression of cancer is the subject of discussion. Hence, the topic of this review is especially attractive to collect the existing endeavors to characterize the viscoelastic features of tumor extracellular matrices and to briefly highlight the present frontiers in cancer progression and escape of cancers from therapy. Finally, this review article illustrates the importance of the tumor extracellular matrix mechano-phenotype, including the phenomenon viscoelasticity in identifying, characterizing, and treating specific cancer types.

Electrospinning and Additive Manufacturing: Adding Three-Dimensionality to Electrospun Scaffolds for Tissue Engineering

The final biochemical and mechanical performance of an implant or scaffold are defined by its structure, as well as the raw materials and processing conditions used during its fabrication. Electrospinning and Additive Manufacturing (AM) are two contrasting processing technologies that have gained popularity amongst the fields of medical research i.e., tissue engineering, implant design, drug delivery. Electrospinning technology is favored for its ability to produce micro- to nanometer fibers from polymer solutions and melts, of which, the dimensions, alignment, porosity, and chemical composition are easily manipulatable to the desired application. AM, on the other hand, offers unrivalled levels of geometrical freedom, allowing highly complex components (i.e., patient-specific) to be built inexpensively within 24 hours. Hence, adopting both technologies together appears to be a progressive step in pursuit of scaffolds that better match the natural architecture of human tissues. Here, we present recent insights into the advances on hybrid scaffolds produced by combining electrospinning (melt electrospinning excluded) and AM, specifically multi-layered architectures consisting of alternating fibers and AM elements, and bioinks reinforced with fibers prior to AM. We discuss how cellular behavior (attachment, migration, and differentiation) is influenced by the co-existence of these micro- and nano-features.

Morphometry of cellular behavior of coelomocytes from starfish Asterias amurensis

A comprehensive statistical analysis using a wide range of linear and non-linear morphological parameters enabled identification of the main stages in the in vitro dynamics of cell behavior of immune cells of the marine invertebrate Asterias amurensis (Echinodermata, Asteroidea). Three stages may be distinguished in the cell behavior, which are characterized by the differences in complexity of the cell boundary microsculpture as well as by the size and asymmetry of the cell and convex hull of the cell. The first stage (5 min after placing cells onto a substrate) is characterized by more complex cell morphology and an increase in the process number and spreading area. The second stage (15 min) is characterized by simplification of cell morphology, retraction of some processes, and rounding of cells upon continued cell spreading. At the third stage (60 min), new large processes with rounded contours emerge due to partial retraction of the flattened cell surface. Each stage is characterized by statistically significant differences in several linear and nonlinear parameters of the external morphology for all cell types.

Effectiveness of Biofunctionalization of Titanium Surfaces with Phosphonic Acid

Surface functionalization of dental implant surfaces has been a developing field in biomaterial research. This study aimed to obtain self-assembled monolayers (SAMs) using carboxyethylphosphonic acid on the surface of titanium (Ti) screws, and assessed the surface characteristics, biomechanical, and cellular behavior on the obtained specimens. This study had three groups, i.e., a control (untreated screws), a test group treated with phosphonic acid, and a third group with treated acid and bone morphogenetic protein (BMP-2) for in vitro analysis of cell lines. The assessed parameters included surface wettability, surface characteristics using scanning electron microscopy (SEM), protein immobilization, and cellular behavior of fibroblasts and mesenchymal stem cells of adipose tissue (MSCat cells). For surface wettability, a Welch test was performed to compare the contact angles between control (67 ± 1.83) and test (18.84 ± 0.72) groups, and a difference was observed in the mean measurements, but was not statistically significant. The SEM analysis showed significant surface roughness on the test screws and the cellular behavior of fibroblasts, and MSCat cells were significantly improved in this group, with fibroblasts having a polygonal shape with numerous vesicles and MSCat cells stable and uniformly coating the test Ti surface. Surface biofunctionalization of Ti surfaces with phosphonic acid showed promising results in this study, but remains to be clinically validated for its applications.

Sprouty3, but Not Sprouty1, Expression Is Beneficial for the Malignant Potential of Osteosarcoma Cells

Sprouty proteins are widely accepted modulators of receptor tyrosine kinase-associated pathways and fulfill diversified roles in cancerogenesis dependent on the originating cells. In this study we detected a high expression of Sprouty3 in osteosarcoma-derived cells and addressed the question of whether Sprouty3 and Sprouty1 influence the malignant phenotype of this bone tumor entity. By using adenoviruses, the Sprouty proteins were expressed in two different cell lines and their influence on cellular behavior was assessed. Growth curve analyses and Scratch assays revealed that Sprouty3 accelerates cell proliferation and migration. Additionally, more colonies were grown in Soft agar if the cells express Sprouty3. In parallel, Sprouty1 had no significant effect on the measured endpoints of the study in osteosarcoma-derived cells. The promotion of the tumorigenic capacities in the presence of Sprouty3 coincided with an increased activation of signaling as measured by evaluating the phosphorylation of extracellular signal-regulated kinases (ERKs). Ectopic expression of a mutated Sprouty3 protein, in which the tyrosine necessary for its activation was substituted, resulted in inhibited migration of the treated cells. Our findings identify Sprouty3 as a candidate for a tumor promoter in osteosarcoma.