Biology subjects

Pendyala, G.

Publications and source records attributed to Pendyala, G..

4 recordsLinked to original sources

bioRxiv2026

Delineating the effects of prenatal oxycodone exposure and melatonin treatment on placental and fetal outcomes in pregnant rats

BackgroundPrenatal oxycodone (oxy) exposure has been associated with adverse pregnancy and fetal developmental outcomes. In this study, we assessed whether chronic prenatal oxy exposure impairs placental and fetal growth in rats and if maternal melatonin supplementation would mitigate these effects. MethodsFemale Sprague-Dawley rats received either saline or oxy via oral gavage for 15 days before mating (10-15mg/kg dose escalation) and throughout pregnancy (15mg/kg). From gestational day (GD) 12.5, half of the dams received melatonin (10mg/kg). On GD19.5, maternal and fetal blood, and maternal, placental and fetal tissues were harvested. Placental histomorphometry was assessed and immunohistochemistry for pan-cytokeratin, PCNA, CD34, -SMA, and TUNEL analysis were performed. Maternal and fetal plasma cytokines, angiogenic factors, and pregnancy hormones were measured by ELISA. Anthropometric data were analyzed using general linear mixed models and other outcomes were analyzed using univariate general linear models. ResultsOxy induced fetal growth restriction as evidenced by reduced placental weight, fetal weight, fetal-to-placental weight ratio, crown-rump length, and fetal liver weight. Melatonin also independently reduced some parameters of fetal growth but when administered with oxy it partially improved fetal outcomes including the head-to-abdominal diameter ratio. Oxy exposure increased placental labyrinth zone area, the percentage of CD34-positive cells, and maternal plasma IL-1{beta} and IL-10 concentrations and reduced the percentage of pan-cytokeratin positive cells, while both oxy and melatonin reduced maternal plasma chorionic gonadotropin levels. ConclusionPrenatal oxy exposure disrupts placental structure, labyrinth anatomy, and induces maternal systemic inflammation, associated with impaired fetal growth. The protective effects of melatonin are partial but indicate a potential brain sparing effect.

developmental biology↗

bioRxiv2025

Prolonged autophagy induction correlates with host cell protein reduction in CHO cell culture

Autophagy, a cellular recycling process regulated by the CLEAR signaling pathway, plays a pivotal role in maintaining cellular homeostasis. We hypothesize that this process may regulate and reduce HCP levels by targeting intracellular proteins and organelles for degradation. This study investigates the relationship between autophagy induction and the reduction of high-risk host cell proteins (HCPs), including polysorbate-degrading enzymes (PSDEs), to enhance the stability of therapeutic biologics such as monoclonal antibodies (mAbs). Using clonal analysis, we identified upregulation of the CLEAR pathway in one clone, correlating with a significant reduction in lipase activity and PSDE abundance. Furthermore, autophagy modulators, such as 3-methyladenine (3-MA), selectively decreased PSDE levels in both batch and fed-batch cultures. This resulted in 62% reduction in lipase activity that corresponded to a 22% improvement in polysorbate-80 stability. Additionally, 3-MA treatment increased mAb specific productivity and altered glycosylation profiles, increasing afucosylation and galactosylation levels. These findings highlight autophagy induction as a promising strategy to modulate product quality profiles and reduce high-risk HCPs in biologics production.

bioengineering↗

bioRxiv2025

Discovery of Chemical Tools for Polysorbate-Degradative Enzyme Control in Biopharmaceutical Upstream Process via Multi-Omic Profiling of Host Cell Clones

Host cell proteins are process-related impurities in biotherapeutics and can potentially pose risks to patient safety and product quality. Specifically, certain host cell-derived enzymes, including lipases, can degrade the formulation excipient polysorbate (PS) in biopharmaceutical formulations, affecting drug product stability in liquid formulations. We leveraged multi-omics approaches, including transcriptomics, proteomics, and activity-based protein profiling (ABPP), to identify mechanisms that regulate PS-degradative enzyme (PSDE) abundance and to develop strategies for their control. Comparative multiomics analysis of two monoclonal antibodies (mAb)-producing host cell clones revealed differential lipase profiles at the mRNA, protein, and enzyme activity levels and associated increased lipase activity with upregulated lipid catabolic pathways such as the fatty acid beta oxidation pathway. Further, for the first time in the literature, we identified peroxisome proliferator-activated receptor {gamma} (PPAR{gamma}) as a key regulator of PSDEs in manufacturing Chinese Hamster Ovary (CHO) cells. Downregulation of the PPAR{gamma} pathway with its antagonists resulted in selective reduction of PSDE levels and improved PS stability without compromising mAb productivity or quality. This study highlights the potential of PPAR{gamma} modulators as chemical tools for PSDE control at the gene regulation level, offering significant implications for biopharmaceutical process development and control.

bioengineering↗

bioRxiv2024

Microglia respond to elevated intraocular pressure and synapse loss in the visual thalamus in a mouse model of glaucoma.

Microglia are resident immune cells of the central nervous system and mediate a broad array of adaptations and responses during disease, injury, and development. Typically, microglia morphology is understood to provide a window into their functional state. However, it is apparent that they have the capacity to adopt a broad spectrum of functional phenotypes characterized by numerous morphological profiles and associated gene expression profiles. Glaucoma, which leads to blindness from retinal ganglion cell (RGC) degeneration, is commonly associated with elevated intraocular pressure and has been shown to trigger microglia responses within the retinal layers, at the optic nerve head, and in retinal projection targets in the brain. The goal of this study was to determine the relationship of microglia morphology to intraocular pressure and the loss of retinal ganglion cell output synapses in the dorsolateral geniculate nucleus (dLGN), a RGC projection target in the thalamus that conveys information to the primary visual cortex. We accomplished this by analyzing dLGN microglia morphologies in histological sections from DBA/2J mice, which develop a form of inherited glaucoma. Microglia morphology was analyzed using skeletonized Iba1-fluorescence images and fractal analyses of individually reconstructed microglia cells. We found that microglia adopted more simplified morphologies, characterized by fewer endpoints and less total process length per microglia cell. There was an age-dependent shift in microglia morphology in tissue from control mice (DBA/2JGpnmb+) that was accelerated in DBA/2J mice. Microglia morphological measurements correlated with cumulative intraocular pressure, immunofluorescence labeling for the complement protein C1q, and density of vGlut2-labeled RGC axon terminals. Additionally, fractal analysis revealed a clear distinction between control and glaucoma dLGN, with microglia from ocular hypertensive DBA/2J dLGN tissue showing an elongated rod-like morphology. RNA-sequencing of dLGN tissue samples showed an upregulation of immune system-related gene expression and several specific genes associated with microglia activation and potential neuroprotective functions. These results suggest that microglia in the dLGN alter their physiology to respond to RGC degeneration in glaucoma, potentially contributing to CNS adaptations to neurodegenerative vision loss.

neuroscience↗