A spatiotemporal proteomic map of human adipogenesis.

White adipocytes function as major energy reservoirs in humans by storing substantial amounts of triglycerides, and their dysfunction is associated with metabolic disorders; however, the mechanisms underlying cellular specialization during adipogenesis remain unknown. Here, we generate a spatiotemporal proteomic atlas of human adipogenesis, which elucidates cellular remodelling as well as the spatial reorganization of metabolic pathways to optimize cells for lipid accumulation and highlights the coordinated regulation of protein localization and abundance during adipocyte formation. We identify compartment-specific regulation of protein levels and localization changes of metabolic enzymes to reprogramme branched-chain amino acids and one-carbon metabolism to provide building blocks and reduction equivalents. Additionally, we identify C19orf12 as a differentiation-induced adipocyte lipid droplet protein that interacts with the translocase of the outer membrane complex of lipid droplet-associated mitochondria and regulates adipocyte lipid storage by determining the capacity of mitochondria to metabolize fatty acids. Overall, our study provides a comprehensive resource for understanding human adipogenesis and for future discoveries in the field.

Deletion of Carboxypeptidase E in ß-Cells Disrupts Proinsulin Processing but Does Not Lead to Spontaneous Development of Diabetes in Mice.

Carboxypeptidase E (CPE) facilitates the conversion of prohormones into mature hormones and is highly expressed in multiple neuroendocrine tissues. Carriers of CPE mutations have elevated plasma proinsulin and develop severe obesity and hyperglycemia. We aimed to determine whether loss of Cpe in pancreatic ß-cells disrupts proinsulin processing and accelerates development of diabetes and obesity in mice. Pancreatic ß-cell-specific Cpe knockout mice (ßCpeKO; Cpefl/fl x Ins1Cre/+) lack mature insulin granules and have elevated proinsulin in plasma; however, glucose-and KCl-stimulated insulin secretion in ßCpeKO islets remained intact. High-fat diet-fed ßCpeKO mice showed weight gain and glucose tolerance comparable with those of Wt littermates. Notably, ß-cell area was increased in chow-fed ßCpeKO mice and ß-cell replication was elevated in ßCpeKO islets. Transcriptomic analysis of ßCpeKO ß-cells revealed elevated glycolysis and Hif1α-target gene expression. On high glucose challenge, ß-cells from ßCpeKO mice showed reduced mitochondrial membrane potential, increased reactive oxygen species, reduced MafA, and elevated Aldh1a3 transcript levels. Following multiple low-dose streptozotocin injections, ßCpeKO mice had accelerated development of hyperglycemia with reduced ß-cell insulin and Glut2 expression. These findings suggest that Cpe and proper proinsulin processing are critical in maintaining ß-cell function during the development of hyperglycemia. ARTICLE HIGHLIGHTS: Carboxypeptidase E (Cpe) is an enzyme that removes the carboxy-terminal arginine and lysine residues from peptide precursors. Mutations in CPE lead to obesity and type 2 diabetes in humans, and whole-body Cpe knockout or mutant mice are obese and hyperglycemic and fail to convert proinsulin to insulin. We show that ß-cell-specific Cpe deletion in mice (ßCpeKO) does not lead to the development of obesity or hyperglycemia, even after prolonged high-fat diet treatment. However, ß-cell proliferation rate and ß-cell area are increased, and the development of hyperglycemia induced by multiple low-dose streptozotocin injections is accelerated in ßCpeKO mice.

Islet amyloid polypeptide does not suppress pancreatic cancer.

OBJECTIVES: Pancreatic cancer risk is elevated approximately two-fold in type 1 and type 2 diabetes. Islet amyloid polypeptide (IAPP) is an abundant beta-cell peptide hormone that declines with diabetes progression. IAPP has been reported to act as a tumour-suppressor in p53-deficient cancers capable of regressing tumour volumes. Given the decline of IAPP during diabetes development, we investigated the actions of IAPP in pancreatic ductal adenocarcinoma (PDAC; the most common form of pancreatic cancer) to determine if IAPP loss in diabetes may increase the risk of pancreatic cancer. METHODS: PANC-1, MIA PaCa-2, and H1299 cells were treated with rodent IAPP, and the IAPP analogs pramlintide and davalintide, and assayed for changes in proliferation, death, and glycolysis. An IAPP-deficient mouse model of PDAC (Iapp-/-; Kras+/LSL-G12D; Trp53flox/flox; Ptf1a+/CreER) was generated for survival analysis. RESULTS: IAPP did not impact glycolysis in MIA PaCa-2 cells, and did not impact cell death, proliferation, or glycolysis in PANC-1 cells or in H1299 cells, which were previously reported as IAPP-sensitive. Iapp deletion in Kras+/LSL-G12D; Trp53flox/flox; Ptf1a+/CreER mice had no effect on survival time to lethal tumour burden. CONCLUSIONS: In contrast to previous reports, we find that IAPP does not function as a tumour suppressor. This suggests that loss of IAPP signalling likely does not increase the risk of pancreatic cancer in individuals with diabetes.

The ß Cell in Diabetes: Integrating Biomarkers With Functional Measures.

The pathogenesis of hyperglycemia observed in most forms of diabetes is intimately tied to the islet ß cell. Impairments in propeptide processing and secretory function, along with the loss of these vital cells, is demonstrable not only in those in whom the diagnosis is established but typically also in individuals who are at increased risk of developing the disease. Biomarkers are used to inform on the state of a biological process, pathological condition, or response to an intervention and are increasingly being used for predicting, diagnosing, and prognosticating disease. They are also proving to be of use in the different forms of diabetes in both research and clinical settings. This review focuses on the ß cell, addressing the potential utility of genetic markers, circulating molecules, immune cell phenotyping, and imaging approaches as biomarkers of cellular function and loss of this critical cell. Further, we consider how these biomarkers complement the more long-established, dynamic, and often complex measurements of ß-cell secretory function that themselves could be considered biomarkers.

Rapid prototyping of custom radiocontrast agent markers for computed tomography-guided procedures.

The objective of this study was to evaluate a method for printing a custom radiocontrast agent mixture to develop computed tomography markers of various shapes and sizes for assisting physicians in computed tomography-guided procedures. The radiocontrast agent mixture was designed to be bright in a computed tomography image, able to be extruded from a nozzle as a liquid and transition into a solid, and sufficiently viscous to be extruded through the tip of a needle in a controlled manner. A mixture printing method was developed using a syringe to house the mixture, a syringe pump to extrude the mixture, and a computer numeric control laser cutter to direct the nozzle in the desired path. To assess the efficacy of printing the radiocontrast agent mixture, we printed several designs, collected computed tomography images, and evaluated various physical properties of the printing method and the resulting computed tomography markers. The average line thickness was 1.56 mm (standard deviation of 0.19 mm, n = 30), the infill percentage was 99.9%, and the deviation in roundness was 0.23 mm (n = 30). These results demonstrated the ability of the proposed method to create various types of skin markers, such as dots, lines, and hollow or solid shapes. Additionally, flat printed patterns can be folded to form three-dimensional structures that can be used to guide and support needle insertions.

Origami Lesion-Targeting Device for CT-Guided Interventions.

The objective of this study is to preliminarily evaluate a lesion-targeting device for CT-guided interventions. The device is created by laser cutting the structure from a sheet of medical grade paperboard, 3D printing two radiocontrast agent grids onto the surface and folding the structure into a rectangular prism with a viewing window. An abdominal imaging phantom was used to evaluate the device through CT imaging and the targeting of lesions for needle insertion. The lesion-targeting trials resulted in a mean targeting error of 2.53 mm (SD 0.59 mm, n = 30). The device is rigid enough to adequately support standard biopsy needles, and it attaches to the patient, reducing the risk of tissue laceration by needles held rigidly in place by an external manipulator. Additional advantages include adequate support for the insertion of multiple surgical tools at once for procedures such as composite ablation and the potential to guide off-axial needle insertion. The low-cost and disposability of the device make it well-suited for the minimally invasive image-guided therapy environment.

MR-Conditional SMA-Based Origami Joint.

Foldable origami structures have been implemented into robotics as a way of compacting joints and circuitry into smaller structures. This technique is especially useful in minimally invasive surgical instruments, where the goal is to create slimline devices that can be inserted through small incisions. Origami also has the potential to cut costs by reducing the amount of material required for assembly. Origami devices are especially suitable for MRI-guided procedures, where instruments must be nonmagnetic because origami is more suitable for flexible, non-metallic materials. MR conditional surgical instruments enable intraoperative MRI procedures that provide superior imaging capabilities to physicians to allow for safer procedures. This work presents an MR conditional joint developed using origami techniques that reduces costs by eliminating assembly of various components and has potential applications in endoscopy. The joint is a compliant rolling-contact element that employs curved-folding origami techniques. A chain of these joints can be constructed from a single sheet of material, eliminating assembly of numerous materials to produce a final product, which is specifically advantageous for constructing low-cost, disposable surgical devices. The prototype contains a degree of bending of ±9 degrees per joint, a response time of less than 4 seconds and an actuation force of 0.5 N using a 1.25 A current. The MRI results showed a minimal artifact of less than 1 mm measured from the boundary of the joint chain and a SNR reduction of less than 10%.

Islet prohormone processing in health and disease.

Biosynthesis of peptide hormones by pancreatic islet endocrine cells is a tightly orchestrated process that is critical for metabolic homeostasis. Like neuroendocrine peptides, insulin and other islet hormones are first synthesized as larger precursor molecules that are processed to their mature secreted products through a series of proteolytic cleavages, mediated by the prohormone convertases Pc1/3 and Pc2, and carboxypeptidase E. Additional posttranslational modifications including C-terminal amidation of the ß-cell peptide islet amyloid polypeptide (IAPP) by peptidyl-glycine α-amidating monooxygenase (Pam) may also occur. Genome-wide association studies (GWAS) have showed genetic linkage of these processing enzymes to obesity, ß-cell dysfunction, and type 2 diabetes (T2D), pointing to their important roles in metabolism and blood glucose regulation. In both type 1 diabetes (T1D) and T2D, and in the face of metabolic or inflammatory stresses, islet prohormone processing may become impaired; indeed elevated proinsulin:insulin (PI:I) ratios are a hallmark of the ß-cell dysfunction in T2D. Recent studies suggest that genetic or acquired defects in proIAPP processing may lead to the production and secretion of incompletely processed forms of proIAPP that could contribute to T2D pathogenesis, and additionally that impaired processing of both PI and proIAPP may be characteristic of ß-cell dysfunction in T1D. In islet α-cells, the prohormone proglucagon is normally processed to bioactive glucagon by Pc2 but may express Pc1/3 under certain conditions leading to production of GLP-1(7-36NH2 ). A better understanding of how ß-cell processing of PI and proIAPP, as well as α-cell processing of proglucagon, are impacted by genetic susceptibility and in the face of diabetogenic stresses, may lead to new therapeutic approaches for improving islet function in diabetes.

Modular force approximating soft robotic pneumatic actuator.

PURPOSE: Soft robots are highly flexible and adaptable instruments that have proven extremely useful, especially in the surgical environment where compliance allows for improved maneuverability throughout the body. Endoscopic devices are a primary example of an instrument that physicians use to navigate to difficult-to-reach areas inside the body. This paper presents a modular soft robotic pneumatic actuator as a proof of concept for a compliant endoscopic device. METHODS: The actuator is 3D printed using an FDM printer. Maximum bending angle is measured using image processing in MATLAB at a gauge pressure level of 35 psi. End-effector displacement is measured using electromagnetic tracking as gauge pressure ranges from 10 to 35 psi, and uniaxial tensile loading ranges from 0 to 120 g. RESULTS: The actuator achieves a maximum bending angle of 145°. Fourth-order polynomial regression is used to model the actuator displacement upon inflation and tensile loading with an average coefficient of correlation value of 0.998. We also develop a feedforward neural network as a robust computer-assisted method for controlling the actuator that achieves a coefficient of correlation value of 0.996. CONCLUSION: We propose a novel modular soft robotic pneumatic actuator that is developed via rapid prototyping and evaluated using image processing and machine learning models. The curled resting shape allows for simple manufacturing and achieves a greater range of bending than other actuators of its kind. A feedforward neural network provides accurate prediction of end-effector displacement upon inflation and loading to deliver precise manipulation and control.