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Mitochondrial architecture has long been associated with metabolic flexibility, although the precise causalities and the underlying mechanisms are still enigmatic. We showed that mitochondrial fragmentation is highly associated with fatty acid oxidation (FAO) rates. Furthermore, forced mitochondrial elongation using Mfn2 over-expression or Drp1 depletion significantly decreased FAO rates, while Mfn2 knockout lead to enhanced FAO rates, suggesting that mitochondrial fragmentation is functionally coupled with fatty acid utilization. Notably, the increased FAO rates upon mitochondrial fragmentation is attributed to decreased CPT1 sensitivity to malonyl-CoA, the endogenous CPT1 inhibitor, consistent with the observation that mitochondrial fragmentation specifically facilitates long chain-fatty acid oxidation (LCFAO), but not short chain fatty acid oxidation. Furthermore, we show pharmacological activators of CPT1 enhance FAO of elongate dmitochondria but not in the presence of malonyl-CoA binding. Suggesting mitochondrial morphology influences CPT1’s binding to a CoA moiety. Taken together, our study provides a biochemical and mechanistic explanation of how mitochondrial architecture links cellular metabolic needs to differential fuel utilization, such as LCFAO, which may be implicated in a myriad of human physiologies and pathologies. In non-alcoholic steatohepatitis (NASH), increased Drp1 mediated mitochondrial fragmentation has been observed in hepatocytes. NASH is characterized by liver inflammation, increased hepatocyte swelling, and some degree of fibrosis. Hepatic steatosis can occur when glucose and fatty acid availability exceeds the energy demand of the liver. This presents the question, is increased fission in NASH a compensatory or a pathogenic mechanism? To address this question, we decreased Drp1 function in liver from adult mice with established NASH (26 week-GAN diet) using GalNAc-siRNA mediated delivery. While NASH alone only elevated circulating AST an

Background: CD44 is a multifunctional membrane glycoprotein. Through its heparan sulfate chain, CD44 presents growth factors to their receptors. We have shown that CD44 and Tropomyosin kinase A (TrkA) form a complex following nerve growth factor (NGF) induction. Our study aimed to understand how CD44 and TrkA interact and the consequences of inhibiting this interaction regarding the pro-tumoral effect of NGF in breast cancer. Methods: After determining which CD44 isoforms (variants) are involved in forming the TrkA/CD44 complex using proximity ligation assays, we investigated the molecular determinants of this interaction. By molecular modeling, we isolated the amino acids involved and confirmed their involvement using mutations. A CD44v3 mimetic peptide was then synthesized to block the TrkA/CD44v3 interaction. The effects of this peptide on the growth, migration and invasion of xenografted triple-negative breast cancer cells were assessed. Finally, we investigated the correlations between the expression of the TrkA/CD44v3 complex in tumors and histo-pronostic parameters. Results: We demonstrated that isoform v3 (CD44v3), but not v6, binds to TrkA in response to NGF stimulation. The final 10 amino acids of exon v3 and the TrkA H112 residue are necessary for the association of CD44v3 with TrkA. Functionally, the CD44v3 mimetic peptide impairs not only NGF-induced RhoA activation, clonogenicity, and migration/invasion of breast cancer cells in vitro but also tumor growth and metastasis in a xenograft mouse model. We also detected TrkA/CD44v3 only in cancerous cells, not in normal adjacent tissues.

Ciliary neurotrophic factor (CNTF) acts as a potent neuroprotective cytokine in multiple models of retinal degeneration. To understand mechanisms underlying its broad neuroprotective effects, we have investigated the influence of CNTF on metabolism in a mouse model of photoreceptor degeneration. CNTF treatment improves the morphology of photoreceptor mitochondria, but also leads to reduced oxygen consumption and suppressed respiratory chain activities. Molecular analyses show elevated glycolytic pathway gene transcripts and active enzymes. Metabolomics analyses detect significantly higher levels of ATP and the energy currency phosphocreatine, elevated glycolytic pathway metabolites, increased TCA cycle metabolites, lipid biosynthetic pathway intermediates, nucleotides, and amino acids. Moreover, CNTF treatment restores the key antioxidant glutathione to the wild type level. Therefore, CNTF significantly impacts the metabolic status of degenerating retinas by promoting aerobic glycolysis and augmenting anabolic activities. These findings reveal cellular mechanisms underlying enhanced neuronal viability and suggest potential therapies for treating retinal degeneration.

In type 1 diabetes, dysfunctional glucose regulation occurs due to the death of insulin-producing beta-cells in the pancreatic islets. Initiation of this process is caused by the inheritance of an adaptive immune system that is predisposed to responding to beta-cell antigens, most notably to insulin itself, coupled with unknown environmental insults priming the autoimmune reaction. While autoimmunity is a primary driver in beta-cell death, there is growing evidence that cellular stress participates in the loss of beta-cells. In the beta-cell fragility model, partial loss of islet mass requires compensatory upregulation of insulin production in the remaining islets, driving a cellular stress capable of triggering apoptosis in the remaining cells. The Glis3-Manf axis has been identified as being pivotal to the relative fragility or robustness of stressed islets, potentially operating in both type 1 and type 2 diabetes. Here, we have used an AAV-based gene delivery system to enhance the expression of the anti-apoptotic protein Manf in the beta-cells of NOD mice. Gene delivery substantially lowered the rate of diabetes development in treated mice. Manf-treated mice demonstrated minimal insulitis and superior preservation of insulin production. Our results demonstrating the therapeutic potential of Manf delivery to enhance beta-cell robustness and avert clinical diabetes.

ObjectiveThe contribution of beta-cell dysfunction to type 2 diabetes (T2D) is not restricted to insulinopenia in the late stages of the disease. Elevated fasting insulinemia in normoglycemic humans is a major factor predicting the onset of insulin resistance and T2D, demonstrating an early alteration of beta-cell function in T2D. Moreover, an early and chronic increase in fasting insulinemia contributes to insulin resistance in high-fat diet (HFD)-fed mice. However, whether there are genetic factors that promote beta-cell-initiated insulin resistance remains undefined. Human variants of the mitochondrial transporter ABCB10, which regulates redox by increasing bilirubin synthesis, have been associated with an elevated risk of T2D. The effects of T2D ABCB10 variants on ABCB10 expression and the actions of ABCB10 in beta-cells are unknown.MethodsThe expression of beta-cell ABCB10 was analyzed in published transcriptome datasets from human beta-cells carrying the T2D-risk ABCB10 variant. Insulin sensitivity, beta-cell proliferation, and secretory function were measured in beta-cell-specific ABCB10 KO mice (Ins1Cre-Abcb10flox/flox). The short-term role of beta-cell ABCB10 activity on glucose-stimulated insulin secretion (GSIS) was determined in isolated islets.ResultsCarrying the T2Drisk allele G of ABCB10 rs348330 variant was associated with increased ABCB10 expression in human beta-cells. Constitutive deletion of Abcb10 in beta-cells protected mice from hyperinsulinemia and insulin resistance by limiting HFD-induced beta-cell expansion. An early limitation in GSIS and H2O2-mediated signaling caused by elevated ABCB10 activity can initiate an over-compensatory expansion of beta-cell mass in response to HFD. Accordingly, increasing ABCB10 expression was sufficient to limit GSIS capacity. In health, ABCB10 protein was decreased during islet maturation, with maturation restricting beta-cell proliferation and elevating GSIS. Finally, ex-vivo and short-term deletion of ABCB

Obesity results from an imbalance in energy homeostasis, whereby excessive energy intake exceeds caloric expenditure. Energy can be dissipated out of an organism by producing heat (thermogenesis), explaining the long-standing interest in exploiting thermogenic processes to counteract obesity. Mitochondrial uncoupling is a process that expends energy by oxidizing nutrients to produce heat, instead of ATP synthesis. Energy can also be dissipated through mechanisms that do not involve mitochondrial uncoupling. Such mechanisms include futile cycles described as metabolic reactions that consume ATP to produce a product from a substrate but then converting the product back into the original substrate, releasing the energy as heat. Energy dissipation driven by cellular ATP demand can be regulated by adjusting the speed and number of futile cycles. Energy consuming futile cycles that are reviewed here are lipolysis/fatty acid re-esterification cycle, creatine/phosphocreatine cycle, and the SERCA-mediated calcium import and export cycle. Their reliance on ATP emphasizes that mitochondrial oxidative function coupled to ATP synthesis, and not just uncoupling, can play a role in thermogenic energy dissipation. Here, we review ATP consuming futile cycles, the evidence for their function in humans, and their potential employment as a strategy to dissipate energy and counteract obesity.

Reliability is an inherent challenge for the emerging nonvolatile technology of racetrack memories, and there exists a fundamental relationship between codes designed for racetrack memories and codes with constrained periodicity. Previous works have sought to construct codes that avoid periodicity in windows, yet have either only provided existence proofs or required high redundancy. This paper provides the first constructions for avoiding periodicity that are both efficient (average-linear time) and with low redundancy (near the lower bound). The proposed algorithms are based on iteratively repairing windows which contain periodicity until all the windows are valid. Intuitively, such algorithms should not converge as there is no monotonic progression; yet, we prove convergence with average-linear time complexity by exploiting subtle properties of the encoder. Overall, we both provide constructions that avoid periodicity in all windows, and we also study the cardinality of such constraints., Comment: Accepted to The International Symposium on Information Theory and Its Applications (ISITA) 2022

Digital memristive processing-in-memory overcomes the memory wall through a fundamental storage device capable of stateful logic within crossbar arrays. Dynamically dividing the crossbar arrays by adding memristive partitions further increases parallelism, thereby overcoming an inherent trade-off in memristive processing-in-memory. The algorithmic topology of partitions is highly unique, and was recently exploited to accelerate multiplication (11x with 32 partitions) and sorting (14x with 16 partitions). Yet, the physical implementation of memristive partitions, such as the peripheral decoders and the control message, has never been considered and may lead to vast impracticality. This paper overcomes that challenge with several novel techniques, presenting efficient practical designs of memristive partitions. We begin by formalizing the algorithmic properties of memristive partitions into serial, parallel, and semi-parallel operations. Peripheral overhead is addressed via a novel technique of half-gates that enables efficient decoding with negligible overhead. Control overhead is addressed by carefully reducing the operation set of memristive partitions, while resulting in negligible performance impact, by utilizing techniques such as shared indices and pattern generators. Ultimately, these efficient practical solutions, combined with the vast algorithmic potential, may revolutionize digital memristive processing-in-memory.

Digital processing-in-memory (PIM) architectures are rapidly emerging to overcome the memory-wall bottleneck by integrating logic within memory elements. Such architectures provide vast computational power within the memory itself in the form of parallel bitwise logic operations. We develop novel algorithmic techniques for PIM that, combined with new perspectives on computer arithmetic, extend this bitwise parallelism to the four fundamental arithmetic operations (addition, subtraction, multiplication, and division), for both fixed-point and floating-point numbers, and using both bit-serial and bit-parallel approaches. We propose a state-of-the-art suite of arithmetic algorithms, demonstrating the first algorithm in the literature of digital PIM for a majority of cases - including cases previously considered impossible for digital PIM, such as floating-point addition. Through a case study on memristive PIM, we compare the proposed algorithms to an NVIDIA RTX 3070 GPU and demonstrate significant throughput and energy improvements., Comment: Accepted to IEEE Transactions on Emerging Topics in Computing (TETC)

Aligning the entire genome of an organism is a compute-intensive task. Pre-alignment filters substantially reduce computation complexity by filtering potential alignment locations. The base-count filter successfully removes over 68% of the potential locations through a histogram-based heuristic. This paper presents FiltPIM, an efficient design of the basecount filter that is based on memristive processing-in-memory. The in-memory design reduces CPU-to-memory data transfer and utilizes both intra-crossbar and inter-crossbar memristive stateful-logic parallelism. The reduction in data transfer and the efficient stateful-logic computation together improve filtering time by 100x compared to a CPU implementation of the filter., Comment: Published in 2021 28th IEEE International Conference on Electronics, Circuits, and Systems (ICECS)

Recent research has sought to accelerate cryptographic hash functions as they are at the core of modern cryptography. Traditional designs, however, suffer from the von Neumann bottleneck that originates from the separation of processing and memory units. An emerging solution to overcome this bottleneck is processing-in-memory (PIM): performing logic within the same devices responsible for memory to eliminate data-transfer and simultaneously provide massive computational parallelism. In this paper, we seek to vastly accelerate the state-of-the-art SHA-3 cryptographic function using the memristive memory processing unit (mMPU), a general-purpose memristive PIM architecture. To that end, we propose a novel in-memory algorithm for variable rotation, and utilize an efficient mapping of the SHA-3 state vector for memristive crossbar arrays to efficiently exploit PIM parallelism. We demonstrate a massive energy efficiency of 1,422 Gbps/W, improving a state-of-the-art memristive SHA-3 accelerator (SHINE-2) by 4.6x., Comment: Accepted to International Conference on Modern Circuits and Systems Technologies (MOCAST) 2022

Phenotypic plasticity allows carcinoma cells to transiently acquire the quasi-mesenchymal features necessary to detach from the primary mass and proceed along the invasion-metastasis cascade. A broad spectrum of epigenetic mechanisms is likely to cause the epithelial-to-mesenchymal (EMT) and mesenchymal-to-epithelial (MET) transitions necessary to allow local dissemination and distant metastasis. Here, we report on the role played by alternative splicing (AS) in eliciting phenotypic plasticity in epithelial malignancies with focus on colon cancer. By taking advantage of the coexistence of subpopulations of fully epithelial (EpCAMhi) and quasi-mesenchymal and highly metastatic (EpCAMlo) cells in conventional human cancer cell lines, we here show that the differential expression of ESRP1 and other RNA-binding proteins (RBPs) downstream of the EMT master regulator ZEB1 alters the AS pattern of a broad spectrum of targets including CD44 and NUMB, thus resulting in the generation of specific isoforms functionally associated with increased invasion and metastasis. Additional functional and clinical validation studies indicate that both the newly identified RBPs and the CD44s and NUMB2/4 splicing isoforms promote local invasion and distant metastasis and are associated with poor survival in colon cancer. The systematic elucidation of the spectrum of EMT-related RBPs and AS targets in epithelial cancers, apart from the insights in the mechanisms underlying phenotypic plasticity, will lead to the identification of novel and tumor-specific therapeutic targets.