Table 1 lists a number of putative GPCRs identified by NC-IUPHAR [197], for which preliminary evidence for an endogenous ligand has been published, or for which there exists a potential link to a disease, or disorder. These GPCRs have recently been reviewed in detail [153]. The GPCRs in Table 1 are all Class A, rhodopsin-like GPCRs. Class A orphan GPCRs not listed in Table 1 are putative GPCRs with as-yet unidentified endogenous ligands.Table 1: Class A orphan GPCRs with putative endogenous ligands GPR3GPR4GPR6GPR12GPR15GPR17GPR20 GPR22GPR26GPR31GPR34GPR35GPR37GPR39 GPR50GPR63GPR65GPR68GPR75GPR84GPR87 GPR88GPR132GPR149GPR161GPR183LGR4LGR5 LGR6MAS1MRGPRDMRGPRX1MRGPRX2P2RY10TAAR2 In addition the orphan receptors GPR18, GPR55 and GPR119 which are reported to respond to endogenous agents analogous to the endogenous cannabinoid ligands have been grouped together (GPR18, GPR55 and GPR119).

Amplification of HER2 occurs in 20% of breast cancers. HER2 (known as ErbB2 in mouse) belongs to the Epidermal Growth Factor Receptor (EGFR) family that is responsible for cell growth and survival. While HER2 does not bind ligands, it forms heterodimers with HER1 (also known as EGFR) in the presence of its ligand EGF, and HER4 following stimulation with Neuregulin. In HER2-positive breast cancer where HER2 expression is high, HER2 homodimerizes leading to excessive activation of downstream signalling and cell proliferation. Current HER2-positive breast cancer treatment involves chemotherapy with drugs called anthracyclines, such as Doxorubicin, and an antibody targeting HER2 called Trastuzumab. This combination is effective in killing HER2-positive breast tumours, however, Trastuzumab alone or in combination with anthracyclines leads to severe cardiotoxicity and heart failure in 2.6-11% of cases, making it a dose-limiting side effect. We hypothesise that cardiac toxicity can be mitigated by targeting cardiomyocytes exclusively to express a mutated (but functional) version of HER2 that is not recognised by trastuzumab. We designed 3 HER2 variants wherein the trastuzumab binding domain was mutated to disrupt binding. These HER2 mutants were transfected into HEK293T cells, and we confirmed that all 3 successfully localised to the cell membrane, and did not bind trastuzumab. We have used bioluminescence resonance energy transfer to examine the capacity of all 3 mutants to activate recruitment of Grb2 to the HER2 as part of HER1/HER2 and HER4/HER2 heterodimers; Western blotting also confirming mutants were capable of stimulating ERK and Akt phosphorylation. Future studies will evaluate the effcacy of using AAVs to instruct cardiomyocytes to express these HER2 mutants in a murine model of breast cancer. We predict that this new therapy would not only mitigate cardiac toxicity, but would also permit higher doses of trastuzumab to be used to treat breast cancer. This work is supported by Australia's National Health and Medical Research Council and the Australian Research Council. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Journal Article Acid-sensing ion channel 1a blockade reduces myocardial injury in rodent models of myocardial infarction Get access Meredith A Redd, Meredith A Redd Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, AustraliaCritical Care Research Group, The Prince Charles Hospital Northside Clinical Unit and Faculty of Medicine, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Yusuke Yoshikawa, Yusuke Yoshikawa School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Nemat Khan, Nemat Khan School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Maleeha Waqar, Maleeha Waqar School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Natalie J Saez, Natalie J Saez Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, AustraliaAustralian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, University of Queensland, AustraliaInfensa Bioscience Pty Ltd, Brisbane, QLD 4101, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Jennifer E Outhwaite, Jennifer E Outhwaite Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, AustraliaSchool of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Jake S Russell, Jake S Russell School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Amy D Hanna, Amy D Hanna Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Han S Chiu, Han S Chiu Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, AustraliaInfensa Bioscience Pty Ltd, Brisbane, QLD 4101, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Sing Yan Er, Sing Yan Er Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar ... Show more Neville J Butcher, Neville J Butcher School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Karine Mardon, Karine Mardon Center for Advanced Imaging, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar John F Fraser, John F Fraser Critical Care Research Group, The Prince Charles Hospital Northside Clinical Unit and Faculty of Medicine, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Mark L Smythe, Mark L Smythe Infensa Bioscience Pty Ltd, Brisbane, QLD 4101, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Lachlan D Rash, Lachlan D Rash School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Walter G Thomas, Walter G Thomas School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Glenn F King, Glenn F King Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, AustraliaAustralian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, University of Queensland, AustraliaInfensa Bioscience Pty Ltd, Brisbane, QLD 4101, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Melissa E Reichelt, Melissa E Reichelt School of Biomedical Sciences, University of Queensland, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Nathan J Palpant Nathan J Palpant Institute for Molecular Bioscience, University of Queensland, 306 Carmody Road, St.Lucia, QLD 4072, AustraliaInfensa Bioscience Pty Ltd, Brisbane, QLD 4101, Australia Corresponding author. Email: n.palpant@imb.uq.edu.au https://orcid.org/0000-0002-9334-8107 Search for other works by this author on: Oxford Academic PubMed Google Scholar European Heart Journal, ehad793, https://doi.org/10.1093/eurheartj/ehad793 Published: 14 December 2023 Article history Received: 08 April 2023 Revision received: 24 October 2023 Accepted: 17 November 2023 Published: 14 December 2023

Bitter taste receptors (T2R) are a subfamily of G protein-coupled receptors that enable humans to detect aversive and toxic substances. The ability to discern bitter compounds varies between individuals and is attributed mainly to naturally occurring T2R polymorphisms. T2Rs are also expressed in numerous non-gustatory tissues, including the heart, indicating potential contributions to cardiovascular physiology. In this study. T2Rs that have previously been identified in human cardiac tissues (T2Rs - 10, 14, 30, 31, 46 and 50) and their naturally occurring polymorphisms were functionally characterised. The ligand-dependent signaling responses of some T2R variants were completely abolished (T2R30 Leu252 and T2R46 Met228), whereas other receptor variants had moderate changes in their maximal response, but not potency, relative to wild type. Using a cAMP fluorescent biosensor, we reveal the productive coupling of T2R14, but not the T2R14 Phe201 variant, to endogenous Gαi. Modeling revealed that these variants resulted in altered interactions that generally affected ligand binding (T2R30 Leu252) or Gα protein interactions (T2R46 Met228 and T2R14 Phe201), rather than receptor structural stability. Interestingly, this study is the first to show a difference in signaling for T2R50 Tyr203 (rs1376251) which has been associated with cardiovascular disease. The observation of naturally occurring functional variation in the T2Rs with the greatest expression in the heart is important, as their discovery should prove useful in deciphering the role of T2Rs within the cardiovascular system.

The actions of angiotensin II (Ang II) are mediated by AT1 and AT2 receptors (nomenclature as agreed by the NC-IUPHAR Subcommittee on Angiotensin receptors [63, 155]), which have around 30% sequence similarity. The decapeptide angiotensin I, the octapeptide angiotensin II and the heptapeptide angiotensin III are endogenous ligands. losartan, candesartan, olmesartan, telmisartan, etc. are clinically used AT1 receptor blockers.

Objectives: Black patients have the highest overall incidence rate of early onset colorectal cancer, with many of these patients presenting with more aggressive disease at diagnosis, ultimately leading to decreased overall survival. We aimed to (1) evaluate how race and age affected overall survival in colorectal cancer patients, and (2) determine the different demographic and clinical covariables that may influence survival in younger individuals. Methods: The 2017 National Cancer Database (NCDB) was used to identify all patients that had colorectal cancer between 2004-2017. These patients were then divided into groups according to age (<45 and ≥45 years old) and race (white and black). Overall survival (OS) between white and black groups according to age was compared. Initial testing of survivor functions between groups revealed violations of the proportional hazards assumption. Accordingly, we used parametric maximum likelihood analyses fitting the survivor functions to Weibull distributions. Logistic regression analysis was used to determine univariate and multivariate relationships between the covariates and race for younger subjects. Propensity score matching analysis was also used to control for differences in the demographic or clinical variables between the young black versus white subgroups. Results: Out of 1.4 million potential cases initially identified, 207,823 unique cases were deemed eligible for evaluation based on study criteria. Black patients in the study population were more likely to be female, have medical comorbidities, and come from areas with lower average income and baseline education. OS was lower in older patients of both race categories when compared to the younger cohorts. Among patients older than 45 years, there were no significant differences in proportional hazard of death between black and white patients. However, among those younger than 45 years, younger black patients had significantly increased hazard of death. Regarding disease burden at diagnosis, pathologic characteristics and overall risk of death, there were no significant differences between black and white patients. Conclusions: Overall survival in young black patients with colorectal cancer is significantly reduced when compared to young white patients, even when controlling for demographic and pathologic factors. This suggests that the outcome disparities between black and white patients are complex, and the underlying factors are not well understood.

Phenotypic and transcriptomic evidence of early cardiac aging, and associated mechanisms, were investigated in young to middle-aged male mice (C57Bl/6; ages 8, 16, 32, 48 wks). Left ventricular gene expression (profiled via Illumina MouseWG-6 BeadChips), contractile and coronary function, and stress-resistance were assessed in Langendorff perfused hearts under normoxic conditions and following ischemic insult (20 min global ischemia-45 min reperfusion; I-R). Baseline or normoxic contractile function was unaltered by age, while cardiac and coronary ‘reserves’ (during β-adrenoceptor stimulation; 1 μM isoproterenol) declined by 48 wks. Resistance to I-R injury fell from 16 to 32 wks. Age-dependent transcriptional changes In un-stressed hearts were limited to 104 genes (>1.3-fold; 0.05 FDR), supporting: up-regulated innate defenses (glutathione and xenobiotic metabolism, chemotaxis, interleukins) and catecholamine secretion; and down-regulated extracellular matrix (ECM), growth factor and survival (PI3K/Akt) signaling. In stressed (post-ischemic) myocardium, ∼15-times as many genes (1528) were age-dependent, grouped into 6 clusters (>1.3-fold change; 0.05 FDR): most changing from 16 wks (45 % up/44 % down), a further 5 % declining from 32 wks. Major age-dependent Biological Processes in I-R hearts reveal: declining ATP metabolism, oxidative phosphorylation, cardiac contraction and morphogenesis, phospholipid metabolism and calcineurin signaling; increasing proteolysis and negative control of MAPK; and mixed changes in nuclear transport and angiogenic genes. Pathway analysis supports reductions in: autophagy, stress response, ER protein processing, mRNA surveillance and ribosome/translation genes; with later falls in mitochondrial biogenesis, oxidative phosphorylation and proteasome genes in I-R hearts. Summarizing, early cardiac aging is evident from 16 to 32 wks in male mice, characterized by: declining cardiovascular reserve and stress-resistance, transcriptomic evidence of constitutive stress and altered catecholamine and survival/growth signaling in healthy hearts; and declining stress response, quality control, mitochondrial energy metabolism and cardiac modeling processes in stressed hearts. These very early changes, potentially key substrate for advanced aging, may inform approaches to healthy aging and cardioprotection in the adult heart.

Inhibitor discovery for emerging drug-target proteins is challenging, especially when target structure or active molecules are unknown. Here, we experimentally validate the broad utility of a deep generative framework trained at-scale on protein sequences, small molecules, and their mutual interactions-unbiased toward any specific target. We performed a protein sequence-conditioned sampling on the generative foundation model to design small-molecule inhibitors for two dissimilar targets: the spike protein receptor-binding domain (RBD) and the main protease from SARS-CoV-2. Despite using only the target sequence information during the model inference, micromolar-level inhibition was observed in vitro for two candidates out of four synthesized for each target. The most potent spike RBD inhibitor exhibited activity against several variants in live virus neutralization assays. These results establish that a single, broadly deployable generative foundation model for accelerated inhibitor discovery is effective and efficient, even in the absence of target structure or binder information.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a dense stroma primarily composed of cancer associated fibroblasts (CAFs). These fibroblasts are a diverse population of cells; some groups of fibroblasts have been shown to play tumor supporting roles while others have been shown to be tumor suppressing. Two prolific and well-characterized CAF subtypes—myofibroblasts (myCAFs) and inflammatory CAFs (iCAFs)—could potentially have different regulatory effects on tumor growth. Understanding the tissue origin of these two groups of CAFs could assist in understanding the complex and heterogeneous roles they might play in PDAC development. Previous studies in our group showed that the splanchnic mesenchyme, a layer of mesenchyme adjacent to the fetal pancreatic epithelium, gives rise to the majority of the total CAF population. Here, I aim to test the hypothesis that both iCAFs and myCAFs originate from the splanchnic mesenchyme. myCAFs have been previously observed to be proximal to tumor cells and express αSMA at a higher level, while iCAFs have been observed to be distal to tumor cells and express αSMA at a lower level. Here, we used a genetically engineered mouse PDAC model carrying KrasG12D/+;p53Frt/+;Pdx1FlpO/+;Isl1cre/+;R26Tomato/+ alleles. In this model, epithelial specific FlpO expression leads to Kras activation and p53 deletion, causing tumorigenesis. Simultaneously, splanchnic specific Cre expression leads to permanent tomato expression in the splanchnic descendants. The pancreata of these mice were harvested, fixed, and mounted onto slides. Co-immunostaining with αSMA and tomato was then done. The stained slides were scanned, and the Inform software was used to segment and phenotype the cells. More than 95% of αSMA high cells were tomato positive, and more than 95% of αSMA low cells were also tomato positive. We thus conclude that both myCAFs and iCAFs originate from the splanchnic mesenchyme. Consistent with previous in vitro studies, this lineage tracing study suggests that different CAF subtypes are likely due to patterning influenced by different factors within the tumor microenvironment. Citation Format: Thomas S. Walter, Lu Han, Michael Ostrowski. myCAFs and iCAFs have similar lineage [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1344.

Venoms are excellent model systems for studying evolutionary processes associated with predator-prey interactions. Here, we present the discovery of a peptide toxin, MIITX2-Mg1a, which is a major component of the venom of the Australian giant red bull ant Myrmecia gulosa and has evolved to mimic, both structurally and functionally, vertebrate epidermal growth factor (EGF) peptide hormones. We show that Mg1a is a potent agonist of the mammalian EGF receptor ErbB1, and that intraplantar injection in mice causes long-lasting hypersensitivity of the injected paw. These data reveal a previously undescribed venom mode of action, highlight a role for ErbB receptors in mammalian pain signaling, and provide an example of molecular mimicry driven by defensive selection pressure.

Table 1 lists a number of putative GPCRs identified by NC-IUPHAR [161], for which preliminary evidence for an endogenous ligand has been published, or for which there exists a potential link to a disease, or disorder. These GPCRs have recently been reviewed in detail [121]. The GPCRs in Table 1 are all Class A, rhodopsin-like GPCRs. Class A orphan GPCRs not listed in Table 1 are putative GPCRs with as-yet unidentified endogenous ligands.Table 1: Class A orphan GPCRs with putative endogenous ligands GPR3GPR4GPR6GPR12GPR15GPR17GPR20 GPR22GPR26GPR31GPR34GPR35GPR37GPR39 GPR50GPR63GPR65GPR68GPR75GPR84GPR87 GPR88GPR132GPR149GPR161GPR183LGR4LGR5 LGR6MAS1MRGPRDMRGPRX1MRGPRX2P2RY10TAAR2 In addition the orphan receptors GPR18, GPR55 and GPR119 which are reported to respond to endogenous agents analogous to the endogenous cannabinoid ligands have been grouped together (GPR18, GPR55 and GPR119).

Transforming growth factor (TGF)‐β signalling commences with the engagement of TGF‐β ligand to cell surface TGF‐β receptors (TGFBR) stimulating Smad2 carboxyl‐terminal phosphorylation (phospho‐Smad2C) and downstream biological responses. In several cell models, G protein‐coupled receptors (GPCRs) transactivate the TGF‐β receptors type‐1 (TGFBR1) leading to phospho‐Smad2C, however, we have recently published that in keratinocytes thrombin did not transactivate the TGFBR1. The bulk of TGFBRs reside in the cytosol and in response to protein kinase B (Akt phosphorylation) can translocate to the cell surface increasing the cell's responsiveness to TGF‐β. In this study, we investigate the role of Akt in GPCR transactivation of the TGFBR1. We demonstrate that angiotensin II and thrombin do not phosphorylate Smad2C in human vascular smooth muscle cells and in keratinocytes respectively. We used Akt agonist, SC79 to sensitise the cells to Akt and observed that Ang II and thrombin phosphorylate Smad2C via Akt/AS160‐dependent pathways. We show that SC79 rapidly translocates TGFBRs to the cell surface thus increasing the cell's response to the GPCR agonist. These findings highlight novel mechanistic insight for the role of Akt in GPCR transactivation of the TGFBR1.

The COVID-19 pandemic has highlighted the urgency for developing more efficient molecular discovery pathways. As exhaustive exploration of the vast chemical space is infeasible, discovering novel inhibitor molecules for emerging drug-target proteins is challenging, particularly for targets with unknown structure or ligands. We demonstrate the broad utility of a single deep generative framework toward discovering novel drug-like inhibitor molecules against two distinct SARS-CoV-2 targets — the main protease (Mpro) and the receptor binding domain (RBD) of the spike protein. To perform target-aware design, the framework employs a target sequence-conditioned sampling of novel molecules from a generative model. Micromolar-level in vitro inhibition was observed for two candidates (out of four synthesized) for each target. The most potent spike RBD inhibitor also emerged as a rare non-covalent antiviral with broad-spectrum activity against several SARS-CoV-2 variants in live virus neutralization assays. These results show that a broadly deployable machine intelligence framework can accelerate hit discovery across different emerging drug-targets.

SummaryTerminating the SARS-CoV-2 pandemic relies upon pan-global vaccination. Current vaccines elicit neutralizing antibody responses to the virus spike derived from early isolates. However, new strains have emerged with multiple mutations: P.1 from Brazil, B.1.351 from South Africa and B.1.1.7 from the UK (12, 10 and 9 changes in the spike respectively). All have mutations in the ACE2 binding site with P.1 and B.1.351 having a virtually identical triplet: E484K, K417N/T and N501Y, which we show confer similar increased affinity for ACE2. We show that, surprisingly, P.1 is significantly less resistant to naturally acquired or vaccine induced antibody responses than B.1.351 suggesting that changes outside the RBD impact neutralisation. Monoclonal antibody 222 neutralises all three variants despite interacting with two of the ACE2 binding site mutations, we explain this through structural analysis and use the 222 light chain to largely restore neutralization potency to a major class of public antibodies.

Antibodies are crucial to immune protection against SARS-CoV-2, with some in emergency use as therapeutics. Here, we identify 377 human monoclonal antibodies (mAbs) recognizing the virus spike and focus mainly on 80 that bind the receptor binding domain (RBD). We devise a competition data-driven method to map RBD binding sites. We find that although antibody binding sites are widely dispersed, neutralizing antibody binding is focused, with nearly all highly inhibitory mAbs (IC50 < 0.1 μg/mL) blocking receptor interaction, except for one that binds a unique epitope in the N-terminal domain. Many of these neutralizing mAbs use public V-genes and are close to germline. We dissect the structural basis of recognition for this large panel of antibodies through X-ray crystallography and cryoelectron microscopy of 19 Fab-antigen structures. We find novel binding modes for some potently inhibitory antibodies and demonstrate that strongly neutralizing mAbs protect, prophylactically or therapeutically, in animal models.

The authors would like to point out that the wrong version of Fig. 7 has appeared in the published article. The figure currently appearing as Fig. 7 was, as part of the review process, incorporated into Fig. 6 and a new Fig. 7 was included. The submitted manuscript for review was correct, but unfortunately in the final uploads and proofing an error must have occurred and went undetected. The authors and journal apologise for this error. The correct Fig. 7 is below: [Figure presented]