BACKGROUND:Mixed response, where different lesions within the same patient show discordant responses to treatment, remains poorly understood. To better understand the complex effects of mixed response on patient survival, we devised three different definitions of mixed response. This retrospective analysis provides the first evaluation of the association between mixed response and survival outcomes in patients with rare cancers treated with dual checkpoint blockade using ipilimumab plus nivolumab, based on data from 52 baskets in the DART SWOG S1609 trial. METHODS:We included 438 patients with Response Evaluation Criteria in Solid Tumors (RECIST) V.1.1-measurable disease and at least two target lesions, after exclusions for ineligibility, early death, or missing data. Overall survival (OS) and progression-free survival (PFS) were compared using log-rank tests and Cox regression, stratified by basket and using a day 65 landmark. A mixed response was evaluated using three definitions: Method 1-RECIST discordance across lesions; Method 2-presence of ≥1 lesion with >5 mm increase or ≥1 with >5 mm decrease; and Method 3-same as Method 2 but with a 1 mm cut-off. RESULTS:Mixed response was significantly associated with worse OS and PFS using both Method 1 (OS: HR 1.80; PFS: HR 1.58) and Method 2 (OS: HR 1.55; PFS: HR 1.57) compared with "all SD/stable per lesion". Among patients classified as "SD by RECIST", those who exhibited a mixed per lesion response according to Method 1 had significantly worse OS (median 9.9 months (8.8-12.4)) than those with a non-mixed per lesion response (median 22.7 months (19.2-32.0)). Further stratification showed that any lesion increasing >5 mm was linked to worse outcomes (OS: HR >2.37, p<0.05). CONCLUSIONS:Mixed response was significantly associated with worse survival outcomes in patients treated with dual immune checkpoint inhibitors, even among those with RECIST-defined stable disease. Our findings suggest that the "worst"-responding lesion drives prognosis, underscoring the limitations of RECIST in capturing clinically relevant heterogeneity. This study highlights the need to incorporate lesion-level assessment into immunotherapy decision-making and provides a foundation for guiding earlier transition to the next line therapies or other therapeutic options. Future studies integrating molecular biomarkers are warranted to refine response evaluation criteria and optimize immune checkpoint inhibitor-based strategies.

Abstract Background: KRAS is the most common driver mutation in non-small cell lung cancer (NSCLC), with G12C and G12D among its most frequent point mutations. Patients with KRAS G12D mutations are more likely to be never-smokers with worse response to immune checkpoint inhibitors (ICI) compared to those with KRAS G12C. As KRAS G12D-targeted therapies are under development, a greater understanding of real-world outcomes associated with current standard-of-care treatments is warranted. Methods: We retrospectively analyzed NSCLC patients with and without KRAS mutations in the University of California Health Data Warehouse. Time on treatment (ToT) of first-line ICIs was estimated, and ICI-related overall survival (OS) was defined as the time from the first ICI infusion until death or loss to follow up. Kaplan-Meier analysis, Cox proportional hazards model, and student’s t-test were applied. Results: We identified 3,391 NSCLC patients, including 158 patients (5%) with KRAS G12D, and 735 (22%) with other KRAS mutations. Patients had a median (range) age of 64 (20-89) upon diagnosis and 57% were female (Table). Among 842 patient (25%) who received ICIs, 58% were treated with pembrolizumab, 20% with nivolumab (including 5% with ipilimumab), and 8% with either atezolizumab or durvalumab. Compared to patients with other KRAS mutations, there were more Hispanic patients with KRAS G12D (20 versus 36 patients, p < 0.01). When adjusting for age, race, and ethnicity in a Cox model, patients with KRAS G12D had similar ToT of ICIs (4.3 versus 4.8 months; p = 0.25; HR, 95%CI: 1.30, 0.83-2.04) and ICI-related OS (9.4 versus 14.0 months; p = 0.36; HR, 95%CI: 1.23, 0.79 - 1.92) as those with other KRAS mutations. Conclusion: Harnessing the power of a structured, multicenter data collective, we describe real-world outcomes from patients with KRAS G12D-mutated NSCLC. Citation Format: Jingtong Liang, Tali Azenkot, Sandip Patel. Real world outcomes for patients with KRAS G12D-mutated non-small cell lung cancer: A University of California Health Data Warehouse retrospective analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6637.

Nicotinic acid adenine dinucleotide phosphate (NAADP) is a Ca2+-releasing second messenger that activates two-pore channels (TPCs) on endosomes and lysosomes. Rather than binding TPCs directly, NAADP acts through cytoplasmic NAADP-binding proteins (NAADP-BPs) which are essential for endolysosomal Ca2+ release. Here we characterized the properties of two recombinant, purified NAADP-BPs: Jupiter Microtubule Associated Homolog 2 (JPT2) and like-Sm protein 12 (LSM12). In contrast to LSM12, JPT2 is predicted to be an intrinsically disordered protein, a feature confirmed by circular dichroism and NMR spectroscopy. Under conditions of low Na+ concentration or molecular crowding, JPT2 underwent phase separation, as demonstrated by multiple orthogonal approaches. JPT2 condensates displayed liquid-like behavior and efficiently recruited LSM12, a novel fluorescent NAADP analog, as well as tubulin. JPT2 condensates also interacted with polymerized microtubules and lysosomes isolated from human cell lines. These findings reveal an unexpected capability of NAADP-BPs to undergo phase separation, and segregate with components needed for NAADP-dependent Ca2+ release. We speculate that these signaling condensates dictate cellular NAADP sensitivity, desensitization of NAADP responses, as well as NAADP targeting to TPCs at membrane contact sites between acidic organelles and the endoplasmic reticulum.

Background: In recent years, Infectious bursal disease is continuously occurring even after vaccination in India and requires an inclusive diagnosis. Therefore, the present study was undertaken to diagnose IBD through molecular and culture methods. Methods: One pooled sample, from each of 54 flocks having birds with IBD like symptoms, was collected. History of bird type, age and vaccination was recorded. Samples were subjected to RT-PCR, egg embryo culture and chicken fibroblast cells culture. Result: A total of 49669 out of 517900 (9.59 %) of birds, aging 3-6 weeks, were displaying the signs similar to IBD.In RT-PCR, 21 (38.88%) samples were found positive which belonged to11 (52.38%) vaccinated and 10 (47.62%) unvaccinated flocks.The RT-PCR positive samples were successfully cultivated for the virus through egg embryo and cell culture. The CEF culture was found least sensitive compared to egg embryo culture and RT-PCR.

DPP-4 inhibitors have been shown to reverse amyloid deposition in Alzheimer's disease (AD) patients with cognitive impairment. Ocimum sanctum L. leaves reported the presence of important phytoconstituents which are reported to have DPP-4 inhibitory activity. To investigate the effects of petroleum ether extract of Ocimum sanctum L. (PEOS) in Intracerebroventricular streptozotocin (ICV-STZ) induced AD rats. ICV-STZ (3 mg/kg) was injected bilaterally into male Wistar rats, while sham animals received the artificial CSF. The ICV-STZ-induced rats were administered with three doses of PEOS (100, 200, and 400 mg/kg, p.o.) for thirty days. All experimental rats were subjected to behaviour parameters (radial arm maze task and novel object recognition test), neurochemical parameters such as GLP-1, Aβ42, and TNF-α levels, and histopathological examination (Congo red staining) of the left brain hemisphere. PEOS significantly reversed the spatial learning and memory deficit exhibited by ICV-STZ-induced rats. Furthermore, PEOS also shows promising results in retreating Aβ deposition, TNF α, and increasing GLP-1 levels. The histopathological study also showed a significant dose-dependent reduction in amyloid plaque formation and dense granule in PEOS -treated rats as compared to the ICV-STZ induced rats (Negative control). The results show that extract of Ocimum sanctum L. attenuated ICV-STZ-induced learning and memory deficits in rats and has the potential to be employed in the therapy of AD.

Ion channels possess selectivity filters that are hardwired to ensure the selective passage of ions. Lysosomal two-pore channels are unusual as they are able to switch their cation selectivity in an agonist-specific manner, allowing differential control of organellar activity. TPC2 is permeable to Ca2+ when activated by the calcium-mobilizing messenger NAADP, but largely Na+-selective when activated by the signaling lipid PI(3,5)P2. Co-stimulation increases Ca2+ but not Na+ permeability; however, the molecular basis for these specificity switches is not well understood. Here we show that mutation of TPC2 residues within the distal cytosolic linker, which connects the first voltage-sensing-like domain to the pore, rendered TPC2 largely unable to discriminate its agonists and highly calcium-permeable, even in the presence of PI(3,5)P2. This mutation induced a co-activated-like state by disrupting a network of residues that connects the linker to the activation gate. Such deregulated agonist action increased lysosomal Ca2+ flux and compromised locomotion and viability when expressed in C. elegans. A proximal disease-linked mutation perturbed agonist action in a similar way both in vitro and in vivo. Biased signaling through TPC2 thus proceeds through molecular determinants that are remote from the selectivity filter, affecting Ca2+ permeability, endo-lysosomal integrity and disease. The selectivity of lysosomal TPC2 ion channels for either Ca²⁺ or Na⁺ depends on which agonists activate the channel. This article shows that this plasticity is governed by a distal allosteric network that allows TPC2 to discriminate between agonists and generate specific lysosomal ion fluxes. A distal allosteric network controls agonist-specific ion selectivity.

Endo-lysosomes are considered acidic Ca2+ stores, but direct measurements of luminal Ca2+ within them are limited. Here, we report that the Ca2+-sensitive luminescent protein aequorin does not reconstitute with its cofactor at highly acidic pH but that a significant fraction of the probe is functional within a mildly acidic compartment when targeted to the endo-lysosomal system. We leveraged this probe (ELGA) to report Ca2+ dynamics in this compartment. We show that Ca2+ uptake is ATP-dependent and sensitive to blockers of ER Ca2+ pumps. We find that the Ca2+ mobilizing messenger IP3 evokes robust luminal responses in wild-type cells, but not in IP3R knockout cells. Responses were comparable to those evoked by activation of the endo-lysosomal ion channels TPCs and TRPMLs. Stimulation with IP3-forming agonists also mobilized the store in intact cells. Super-resolution microscopy analysis was consistent with the presence of IP3Rs within the endo-lysosomal system. Our data reveal a physiologically relevant, IP3-sensitive store of Ca2+ within the endo-lysosomal system.

Lung cancer is the leading cause of cancer death in the US and globally. The mortality from lung cancer has been declining, due to a reduction in incidence and advances in treatment. Although recent success in developing targeted and immunotherapies for lung cancer has benefitted patients, it has also expanded the complexity of potential treatment options for health care providers. To aid in reducing such complexity, experts in oncology convened a conference (Bridging the Gaps in Lung Cancer) to identify current knowledge gaps and controversies in the diagnosis, treatment, and outcomes of various lung cancer scenarios, as described here. Such scenarios relate to biomarkers and testing in lung cancer, small cell lung cancer, EGFR mutations and targeted therapy in non-small cell lung cancer (NSCLC), early-stage NSCLC, KRAS/BRAF/MET and other genomic alterations in NSCLC, and immunotherapy in advanced NSCLC.

Electrochemical gradients exist not only across the plasma membrane (PM) but also across membranes of organelles. Various endomembrane-localised ion channels and transporters have been identified, the activity of which is critical for organellar (and also cellular) ionic homeostasis that underpins diverse cellular processes. Aberrant organellar ion flux underlies several diseases, identifying organellar channels and transporters as potential drug targets. Therefore, the need for probing the functions of these proteins in situ cannot be overemphasised. The acidic interior of a few organelles as well as the dynamic nature of most organelles historically presented challenges for reliable estimation of luminal ionic concentrations. But there have been significant methodological and technical advancements by now, allowing measurement of levels of specific ions within these organelles as well as their flux across endomembranes with increasing precision. Evidence also continues to amass reporting mechanosensitivity of the endomembranes and its physiological significance. Here we highlight some recent developments in tools and techniques for measuring the levels and movement of some selected organellar cations as well as organellar mechanosensitivity.

Background Associations between immune-related adverse events from checkpoint inhibitor therapy and outcomes have been previously evaluated, with most prior research finding a positive association between toxicity and survival. This prior research has generally reported on more common tumor types. We use a unique data resource of a federally funded basket trial (NCT02834013) for patients with rare cancers (n = 684) to evaluate associations between immune-related adverse events and overall survival and progression-free survival (PFS). Methods Patients were treated with nivolumab and ipilimumab; the trial was opened at more than 1000 sites. Landmark Cox regression models were used to assess first cycle immune-related adverse event associations with PFS and overall survival. Results We found that grade 1-2 treatment-related immune-related adverse events in the first cycle of therapy were associated with longer overall survival (multivariable hazard ratio [HR] = 0.61, 95% confidence interval [CI] = 0.49 to 0.75; P < .001) compared with no treatment-related immune-related adverse event, while grade 3-4 immune-related adverse events were associated with shorter overall survival (HR = 1.41, 95% CI = 1.04 to 1.90; P = .025). Similar but weaker associations were observed with PFS and grade 1-2 treatment-related immune-related adverse events (HR = 0.83, 95% CI = 0.67 to 1.01; P = .067) and grade 3-4 (HR = 1.35, 95% CI = 1.02 to 1.78; P = .037) compared with no treatment-related immune-related adverse events. Grade 1-2 dermatologic toxicity was associated with improved overall survival compared with other grade 1-2 toxicities (HR = 0.67, 95% CI = 0.52 to 0.85; P = .002). There was no statistically significant overall survival difference between patients with grade 1-2 fatigue, gastrointestinal, metabolic, hepatic, endocrine, and thyroid toxicities vs other grade 1-2 toxicities. Conclusion In this large cohort of patients with rare tumors receiving checkpoint inhibitor therapy, grade of immune-related adverse event in the first cycle was predictive for survival.

Senescence emerged as significant mechanism of aging and age-related diseases, offering an attractive target for clinical interventions. Senescent cells release a senescence-associated secretory phenotype (SASP), including exosomes that may act as signal transducers between distal tissues, and propagate secondary senescence. However, the composition of exosomal SASP components remains underexplored. We identified ~1,300 exosome proteins released by senescent primary human lung fibroblasts induced by three different senescence inducers. In parallel, a small human plasma cohort from young (20-26 years) and old (65-74 years) individuals revealed 1,350 exosome proteins and 171 plasma exosome proteins were altered in old individuals. Of the age-regulated plasma exosome proteins, we observed 52 exosomal SASP factors that were also regulated in exosomes from the senescent fibroblasts, SERPINs, Prothrombin, Coagulation factor V, Plasminogen, and Reelin. We identified 247 exosome lipids. Following senescence induction phosphatidylcholines, phosphatidylethanolamines, and sphingomyelins increased significantly indicating cellular membrane changes. Significantly changed proteins were related to extracellular matrix remodeling and inflammation, both potentially detrimental pathways that can damage surrounding tissues and even induce secondary senescence. Our proof-of-principle study - even though initially from a rather small human cohort - suggested potential senescence biomarker candidates, enabling future surveillance of senescence burden in the aging population.

Parkinson's disease results from degeneration of dopaminergic neurons in the midbrain, but the underlying mechanisms are unclear. Here, we identify novel crosstalk between depolarization-induced entry of Ca2+ and lysosomal cation release in maintaining dopaminergic neuronal function. The common disease-causing G2019S mutation in LRRK2 selectively exaggerated Ca2+ entry in vitro. Chemical and molecular strategies inhibiting the lysosomal ion channel TPC2 reversed this. Using Drosophila, which lack TPCs, we show that the expression of human TPC2 phenocopied LRRK2 G2019S in perturbing dopaminergic-dependent vision and movement in vivo. Mechanistically, dysfunction required an intact pore, correct subcellular targeting and Rab interactivity of TPC2. Reducing Ca2+ permeability with a novel biased TPC2 agonist corrected deviant Ca2+ entry and behavioral defects. Thus, both inhibition and select activation of TPC2 are beneficial. Functional coupling between lysosomal cation release and Ca2+ influx emerges as a potential druggable node in Parkinson's disease.

To grow and divide cells must tightly coordinate anabolic programs with the availability of nutrients and growth factors. This balance is especially critical during postnatal development, when biosynthetic and energetic demands are high, and nutrient supply and neonates have to adapt to periods of fasting. These conditions place acute stress on the proteostasis network, making autophagy essential for nutrient recycling. We found that the chaperone aryl hydrocarbon receptor-interacting protein (AIP) supports both arms of this metabolic balance: promoting anabolic PI3K-AKT signaling for mTORC1 activation and enabling catabolic processes such as proteasomal degradation and autophagy. Loss of AIP causes a severe neonatal metabolic disorder, where affected infants fail to thrive postnatally. Our findings establish AIP as a central regulator of neonatal metabolic adaptation and cellular homeostasis. One Sentence Summary AIP integrates nutrient sensing and protein recycling to sustain neonatal survival. ### Competing Interest Statement The authors have declared no competing interest. Rosetrees Trust, M789 Great Ormond Street Hospital Children's Charity, V4722 China Scholarship Council, https://ror.org/04atp4p48 Clinical Training Fellowship from The Medical College of St Bartholomew’s Trust, MEAG1T1R Medical Research Council, MR/M018539/1 Barts Charity, MGU0549 William Harvey Research Institute, MGU093

The Excellence in Oncology Care (EIOC) 2023 Congress, held in Dubai, took place both in person and online. This annual event brought together oncologists from across the Middle East and the Indian subcontinent. A panel of 17 regional experts worked together to develop the first region-specific consensus guidelines. A survey was conducted among the panelists, followed by a preparatory meeting to discuss the results and formulate the guidelines. These guidelines were then presented and discussed at the congress. The panel addressed non-small cell lung cancer (NSCLC), focusing specifically on integrating immunotherapy in resectable NSCLC. The management of NSCLC is advancing, particularly in the early stages. For stages II to IIIA, adjuvant platinum-based chemotherapy is recommended, but new therapies are needed due to limited efficacy. Immunotherapy, including atezolizumab and pembrolizumab, is now standard for resected stage I-III NSCLC, though challenges in clinical uptake and biomarker testing persist. Comprehensive staging evaluation, including positron emission tomography-computed tomography, mediastinal assessment, and central nervous system screening, is advised. EGFR and ALK biomarker testing, along with multidisciplinary team discussions, is crucial. Adjuvant immunotherapy decisions should be guided by PD-L1 status, with atezolizumab or pembrolizumab recommended for 12 months. Neoadjuvant ICI therapy with chemotherapy is suggested for stages IB to IIIA/B NSCLC. The EIOC guidelines offer detailed insights into NSCLC management.

Podocytopathies, driven by injury to glomerular podocytes, are a major cause of proteinuric kidney diseases like focal segmental glomerulosclerosis (FSGS) and diabetic kidney disease (DKD). The transient receptor potential canonical 6 (TRPC6) channel, expressed in podocytes, is critical for maintaining the glomerular filtration barrier. Dysregulation of TRPC6 activity-whether due to gain-of-function mutations or pathological activation-induces excessive calcium influx, leading to podocyte cytoskeletal disruption, oxidative stress, and apoptosis. While initially linked to hereditary FSGS, emerging evidence implicates TRPC6 dysfunction in diverse acquired podocytopathies, including DKD. This review explores the molecular mechanisms underlying TRPC6-mediated podocyte injury across these conditions, focusing on key calcium-dependent signaling pathways such as calcineurin/NFAT. It critically examines the role of TRPC6 as a therapeutic target, highlighting the rationale for its inhibition and the challenges involved in clinical translation. Understanding these mechanisms is crucial for developing strategies to prevent progression to end-stage renal disease.

Immune checkpoint blockade (ICB) therapies, including anti-CTLA4 (ipilimumab) and anti-PD1 (pembrolizumab), have revolutionized cancer treatment by reinvigorating antitumor immunity. However, ICB therapies often cause immune-related adverse events (irAEs), off-target toxicities limiting clinical utility. Current models, like single humanized CTLA-4 (hCTLA4) knock-in mice, provide insights into ICB biology but fail to fully replicate the multi-organ inflammation seen in patients. Moreover, ipilimumab monotherapy does not mimic the complexity of irAEs from combination therapies. To address these gaps, we developed a double humanized immune checkpoint knock-in (hCTLA4/hPD1) mouse model to study ICB-driven irAEs in solid tumor. This model supports ipilimumab and pembrolizumab, replicating human ICB regimens and enabling evaluation of therapeutic efficacy and adverse effects in the tumor microenvironment. Immunotherapy-prone tumor models were integrated into hCTLA4/hPD1 mice to assess systemic and tumor-specific immune responses to combination ICB. Using this model, we are exploring bioactive lipids, identified by our group, that mitigate ICB-driven irAEs in single hCTLA4 knock-in mice. This double-knock-in model bridges gaps between preclinical ICB models and human irAEs, enabling mechanistic studies of irAEs under clinically relevant regimens. It also facilitates testing interventions like lipid supplementation, advancing safer, more effective immunotherapies. Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)