Strong Inactivation Research Articles

Simultaneous treatment with tap water-based neutral electrolyzed water and citric acid for inactivating foodborne pathogens on stainless steel and their synergistic bactericidal mechanisms

Electrolyzed water has many potential applications, the primary one being the eliminating microorganisms in the food industry. However, this application is limited by high chlorine levels. Tap water-based neutral electrolyzed water (TNEW) has the advantage of relatively low chlorine concentration (1.7–4.6 mg/L), but its sterilization effect is limited. In this study, TNEW was combined with citric acid (CA) solution, resulting in strong synergistic inactivation effect on Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on stainless steel surfaces. Simultaneous treatment of stainless steel with TNEW [available chlorine concentration (ACC) of 4.97 mg/L] and 0.1% CA for 15 min resulted in 4.78-, 4.26-, and >5.69 log CFU/cm2 surface reduction in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, which was a synergistic cell count reductions of 3.25, 2.83, and >3.84 log units, respectively. Fluorescence and RT-qPCR analyses identified synergistic production of intracellular reactive oxygen species (ROS), damage to the cell membrane resulting from lipid peroxidation, and DNA damage as the major factors contributing to the synergistic lethal effect. The results of this study suggest that the TNEW–CA combination treatment can be applied as a potential substitute for conventional electrolyzed water treatment for food contact surface disinfection.

Development of Deep Ultraviolet LED Packaging

Deep UV LED package is a hotspot of growing concern for research scholars, it is through the LED semiconductor light-emitting devices emit UVC band ultraviolet (typical wavelength 260 ~ 280nm), is a new type of healthy artificial light source, compared with the traditional UV light source mercury lamps, the deep UV LED has a wavelength of accurate and controllable, green, and so on many advantages. Due to the high-energy deep UV radiation capability, it has a strong bactericidal and inactivation effect on bacteria, viruses and other microorganisms. In recent years, with the continuous progress of deep packaging technology, the optical efficiency and reliability of deep UV LEDs have been significantly improved. This paper summarizes the key technologies of deep-ultraviolet packaging, the performance of deep-ultraviolet LEDs and the application of deep-ultraviolet LEDs; through the understanding of the packaging materials, packaging structure, packaging process and so on to improve the performance of deep-ultraviolet LEDs, so that it is better to apply to the market, which can be seen that the deep-ultraviolet LEDs have a great prospect for development.

Cold atmospheric plasma inactivates Aspergillus flavus and Fusarium keratoplasticum biofilms and conidia in vitro.

Introduction. Aspergillus flavus and Fusarium keratoplasticum are common causative pathogens of fungal keratitis (FK), a severe corneal disease associated with significant morbidity and vision loss. Escalating incidence of antifungal resistance to available antifungal drugs poses a major challenge to FK treatment. Cold atmospheric plasma (CAP) is a pioneering nonpharmacologic antimicrobial intervention that has demonstrated potential as a broad-spectrum antifungal treatment.Gap statement. Previous research highlights biofilm-associated resistance as a critical barrier to effective FK treatment. Although CAP has shown promise against various fungal infections, its efficacy against biofilm and conidial forms of FK pathogens remains inadequately explored.Aim. This study aims to investigate the antifungal efficacy of CAP against clinical fungal keratitis isolates of A. flavus and F. keratoplasticum in vitro.Methodology. Power parameters (22-27 kVpp, 300-400 Hz and 20-80 mA) of a dielectric barrier discharge CAP device were optimized for inactivation of A. flavus biofilms. Optimal applied voltage and total current were applied to F. keratoplasticum biofilms and conidial suspensions of A. flavus and F. keratoplasticum. The antifungal effect of CAP treatment was investigated by evaluating fungal viability through means of metabolic activity, c.f.u. enumeration (c.f.u. ml-1) and biofilm formation.Results. For both fungal species, CAP exhibited strong time-dependent inactivation, achieving greater than 80 % reduction in metabolic activity and c.f.u. ml-1 within 300 s or less, and complete inhibition after 600 s of treatment.Conclusion. Our findings indicate that CAP is a promising broad-spectrum antifungal intervention. CAP treatment effectively reduces fungal viability in both biofilm and conidial suspension cultures of A. flavus and F. keratoplasticum, suggesting its potential as an alternative treatment strategy for fungal keratitis.

The use of organic peroxyacids for the inactivation of calcium-mediated biofilm formation by Bacillus licheniformis

Biofilms formed by Bacillus licheniformis usually serve as a continuous bacterial contamination in the dairy industry. The purpose of this study was to explore how 1% organic acids (citric acid, phenyllactic acid, malic acid, and butyric acid) and their 1% organic peroxyacids (peroxycitric acid, peroxyphenyllactic acid, peroxymalic acid, and peroxybutyric acid) can inactive the planktonic and biofilm cells of B. licheniformis. Results demonstrated that the number of B. licheniformis planktonic cells was increased from 4 to 6.46 Log CFU mL−1 when treated with water. Organic peroxyacids, especially peroxyphenyllactic acid (PPA), obviously inactivated planktonic cells of B. licheniformis by a range of 3.32–4.14 Log CFU mL−1. Moreover, organic peroxyacids could also decrease the cell surface hydrophobicity of bacteria, and effectively control the Ca2+-mediated biofilm formation by B. licheniformis in 96-well plates (reduction of OD values by 0.14–0.33) and on stainless steel (reduction of cell counts by 1.20–2.49 Log CFU cm−2). The low minimum biofilm eradication concentration of PPA (0.125%) also suggests its strong biofilm inactivation activity, which could serve as a novel and environmentally friendly biofilm control agent in the dairy industry.

Multifunctional interaction of CihC/FbpC orthologs of relapsing fever spirochetes with host-derived proteins involved in adhesion, fibrinolysis, and complement evasion.

Relapsing fever (RF) remains a neglected human disease that is caused by a number of diverse pathogenic Borrelia (B.) species. Characterized by high cell densities in human blood, relapsing fever spirochetes have developed plentiful strategies to avoid recognition by the host defense mechanisms. In this scenario, spirochetal lipoproteins exhibiting multifunctional binding properties in the interaction with host-derived molecules are known to play a key role in adhesion, fibrinolysis and complement activation. Binding of CihC/FbpC orthologs to different human proteins and conversion of protein-bound plasminogen to proteolytic active plasmin were examined by ELISA. To analyze the inhibitory capacity of CihC/FbpC orthologs on complement activation, a microtiter-based approach was performed. Finally, AlphaFold predictions were utilized to identified the complement-interacting residues. Here, we elucidate the binding properties of CihC/FbpC-orthologs from distinct RF spirochetes including B. parkeri, B. hermsii, B. turicatae, and B. recurrentis to human fibronectin, plasminogen, and complement component C1r. All CihC/FbpC-orthologs displayed similar binding properties to fibronectin, plasminogen, and C1r, respectively. Functional studies revealed a dose dependent binding of plasminogen to all borrelial proteins and conversion to active plasmin. The proteolytic activity of plasmin was almost completely abrogated by tranexamic acid, indicating that lysine residues are involved in the interaction with this serine protease. In addition, a strong inactivation capacity toward the classical pathway could be demonstrated for the wild-type CihC/FbpC-orthologs as well as for the C-terminal CihC fragment of B. recurrentis. Pre-incubation of human serum with borrelial molecules except CihC/FbpC variants lacking the C-terminal region protected serum-susceptible Borrelia cells from complement-mediated lysis. Utilizing AlphaFold2 predictions and existing crystal structures, we mapped the putative key residues involved in C1r binding on the CihC/FbpC orthologs attempting to explain the relatively small differences in C1r binding affinity despite the substitutions of key residues. Collectively, our data advance the understanding of the multiple binding properties of structural and functional highly similar molecules of relapsing fever spirochetes proposed to be involved in pathogenesis and virulence.

Photodynamic Eradication of Pseudomonas aeruginosa with Ru-Photosensitizers Encapsulated in Enzyme Degradable Nanocarriers.

The development of new approaches for the treatment of the increasingly antibiotic-resistant pathogen Pseudomonas aeruginosa was targeted by enhancing the effect of local antimicrobial photodynamic therapy (aPDT) using poly(ethylene glycol)-block-poly(lactic acid) (PEG114-block-PLAx) nanocarriers that were loaded with a ruthenium-based photosensitizer (PS). The action of tris(1,10-phenanthroline) ruthenium (II) bis(hexafluorophosphate) (RuPhen3) encapsulated in PEG114-block-PLAx micelles and vesicles was shown to result in an appreciable aPDT inactivation efficiency against planktonic Pseudomonas aeruginosa. In particular, the encapsulation of the PS, its release, and the efficiency of singlet oxygen (1O2) generation upon irradiation with blue light were studied spectroscopically. The antimicrobial effect was analyzed with two strains of Pseudomonas aeruginosa. Compared with PS-loaded micelles, formulations of the PS-loaded vesicles showed 10 times enhanced activity with a strong photodynamic inactivation effect of at least a 4.7 log reduction against both a Pseudomonas aeruginosa lab strain and a clinical isolate collected from the lung of a cystic fibrosis (CF) patient. This work lays the foundation for the targeted eradication of Pseudomonas aeruginosa using aPDT in various medical application areas.

Molecular tuning of sea anemone stinging.

Jellyfish and sea anemones fire single-use, venom-covered barbs to immobilize prey or predators. We previously showed that the anemone Nematostella vectensis uses a specialized voltage-gated calcium (CaV) channel to trigger stinging in response to synergistic prey-derived chemicals and touch (Weir et al., 2020). Here, we use experiments and theory to find that stinging behavior is suited to distinct ecological niches. We find that the burrowing anemone Nematostella uses uniquely strong CaV inactivation for precise control of predatory stinging. In contrast, the related anemone Exaiptasia diaphana inhabits exposed environments to support photosynthetic endosymbionts. Consistent with its niche, Exaiptasia indiscriminately stings for defense and expresses a CaV splice variant that confers weak inactivation. Chimeric analyses reveal that CaVβ subunit adaptations regulate inactivation, suggesting an evolutionary tuning mechanism for stinging behavior. These findings demonstrate how functional specialization of ion channel structure contributes to distinct organismal behavior.

Molecular tuning of sea anemone stinging

Jellyfish and sea anemones fire single-use, venom-covered barbs to immobilize prey or predators. We previously showed that the anemone Nematostella vectensis uses a specialized voltage-gated calcium (CaV) channel to trigger stinging in response to synergistic prey-derived chemicals and touch (Weir et al., 2020). Here, we use experiments and theory to find that stinging behavior is suited to distinct ecological niches. We find that the burrowing anemone Nematostella uses uniquely strong CaV inactivation for precise control of predatory stinging. In contrast, the related anemone Exaiptasia diaphana inhabits exposed environments to support photosynthetic endosymbionts. Consistent with its niche, Exaiptasia indiscriminately stings for defense and expresses a CaV splice variant that confers weak inactivation. Chimeric analyses reveal that CaVβ subunit adaptations regulate inactivation, suggesting an evolutionary tuning mechanism for stinging behavior. These findings demonstrate how functional specialization of ion channel structure contributes to distinct organismal behavior.

Strong Intracellular Signal Inactivation Produces Sharper and More Robust Signaling from Cell Membrane to Nucleus.

Strong Intracellular Signal Inactivation Produces Sharper and More Robust Signaling from Cell Membrane to Nucleus.

Virus purification highlights the high susceptibility of SARS-CoV-2 to a chlorine-based disinfectant, chlorous acid.

Chlorous acid water (HClO2) is known for its antimicrobial activity. In this study, we attempted to accurately assess the ability of chlorous acid water to inactivate SARS-CoV-2. When using cell culture supernatants of infected cells as the test virus, the 99% inactivation concentration (IC99) for the SARS-CoV-2 D614G variant, as well as the Delta and Omicron variants, was approximately 10ppm of free chlorine concentration with a reaction time of 10 minutes. On the other hand, in experiments using a more purified virus, the IC99 of chlorous acid water was 0.41-0.74ppm with a reaction time of 1 minute, showing a strong inactivation capacity over 200 times. With sodium hypochlorite water, the IC99 was 0.54ppm, confirming that these chlorine compounds have a potent inactivation effect against SARS-CoV-2. However, it became clear that when using cell culture supernatants of infected cells as the test virus, the effect is masked by impurities such as amino acids contained therein. Also, when proteins (0.5% polypeptone, or 0.3% BSA + 0.3% sheep red blood cells, or 5% FBS) were added to the purified virus, the IC99 values became high, ranging from 5.3 to 76ppm with a reaction time of 10 minutes, significantly reducing the effect. However, considering that the usual usage concentration is 200ppm, it was shown that chlorous acid water can still exert sufficient disinfection effects even in the presence of proteins. Further research is needed to confirm the practical applications and effects of chlorous acid water, but it has the potential to be an important tool for preventing the spread of SARS-CoV-2.

PDGFβ receptor-targeted delivery of truncated transforming growth factor β receptor type II for improving the in vitro and in vivo anti-renal fibrosis activity via strong inactivation of TGF-β1/Smad signaling pathway.

Truncated transforming growth factor β receptor type II (tTβRII), serving as a trap for binding excessive transforming growth factor β1 (TGF-β1) by means of competing with wild-type TβRII, is a promising strategy for the treatment of kidney fibrosis. Platelet-derived growth factor β receptor (PDGFβR) is highly expressed in interstitial myofibroblasts in kidney fibrosis. This study identified the interaction between a novel tTβRII variant Z-tTβRII (PDGFβR-specific affibody ZPDGFβR fused to the N-terminus of tTβRII) and TGF-β1. Moreover, Z-tTβRII highly targeted to TGF-β1-activated NIH3T3 cells and UUO-induced fibrotic kidney, but less to normal cells, tissues, and organs. Furthermore, Z-tTβRII significantly inhibited cell proliferation and migration, and reduced fibrosis markers expression and phosphorylation level of Smad2/3 in activated NIH3T3 cells. Meanwhile, Z-tTβRII markedly alleviated the kidney histopathology and fibrotic responses, and inhibited the TGF-β1/Smad signaling pathway in UUO mice. Besides, Z-tTβRII showed good safety performance in the treatment of UUO mice. In conclusion, these results demonstrated that Z-tTβRII may be a potential candidate for a targeting therapy on renal fibrosis due to the high potential of fibrotic kidney-targeting and strong anti-renal fibrosis activity.

Physiologically Based Pharmacokinetic Modeling of Bergamottin and 6,7-Dihydroxybergamottin to Describe CYP3A4 Mediated Grapefruit-Drug Interactions.

Grapefruit is a moderate to strong inactivator of CYP3A4, which metabolizes up to 50% of marketed drugs. The inhibitory effect is mainly attributed to furanocoumarins present in the fruit, irreversibly inhibiting preferably intestinal CYP3A4 as suicide inhibitors. Effects on CYP3A4 victim drugs can still be measured up to 24 hours after grapefruit juice (GFJ) consumption. The current study aimed to establish a physiologically-based pharmacokinetic (PBPK) grapefruit-drug interaction model by modeling the relevant CYP3A4 inhibiting ingredients of the fruit to simulate and predict the effect of GFJ consumption on plasma concentration-time profiles of various CYP3A4 victim drugs. The grapefruit model was developed in PK-Sim and coupled with previously developed PBPK models of CYP3A4 substrates that were publicly available and already evaluated for CYP3A4-mediated drug-drug interactions. Overall, 43 clinical studies were used for model development. Models of bergamottin (BGT) and 6,7-dihydroxybergamottin (DHB) as relevant active ingredients in GFJ were established. Both models include: (i) CYP3A4 inactivation informed by in vitro parameters, (ii) a CYP3A4 mediated clearance estimated during model development, as well as (iii) passive glomerular filtration. The final model successfully describes interactions of GFJ ingredients with 10 different CYP3A4 victim drugs, simulating the effect of the CYP3A4 inactivation on the victims' pharmacokinetics as well as their main metabolites. Furthermore, the model sufficiently captures the time-dependent effect of CYP3A4 inactivation as well as the effect of grapefruit ingestion on intestinal and hepatic CYP3A4 concentrations.

Evaluation of Alternative-to-Gas Chlorination Disinfection Technologies in the Treatment of Maltese Potable Water

For years, gas chlorination has been the adopted disinfection technology in the treatment of Maltese potable water. Despite its strong bacterial inactivation potential, traditional chlorination generates high free chlorine residual and disinfection by-products that compromise the sensory attributes of drinking water and deter the population from consuming it. We have identified plausible alternative-to-gas-chlorination technologies for its treatment, with the aim of (a) reducing the disinfectant and/or chlorination dose used for microbial inactivation, and (b) attenuating the negative impact of putative disinfection by-products on the water’s organolepsis, while safeguarding its safe-for-consumption characteristics. We have subjected ultraviolet C (UVC) irradiation, hydrodynamic cavitation (HC), ClO2 generation, and electrochlorination (NaClO) to bacteriological and physicochemical bench-scale studies to assess their bacterial inactivation efficacy and by-product generation propensity, respectively. All the tested technologies except HC achieved a minimum of 3 Log10 microbial inactivation, with NaClO and ClO2 appearing more effective over neutral and alkaline pH conditions, respectively. In addition, we have identified synergistic effects of cavitation on UVC for Enterococcus faecalis inactivation, stemming from enhancement in oxidative stress. Moderate reductions in the total dissolved solid content and Ca2+ hardness of the tested water also emerged following prolonged cavitation. For feasibility studies, the performance of the technologies was further evaluated on the following areas: (a) implementation, (b) practicality, (c) adaptability, (d) integration, (e) environment and sustainability, and (f) cost and effect. Electrochemical generation of NaClO emerged as the most promising technology for further on-site work, followed by ClO2 and UVC.

Antibacterial Mechanism of Shikonin Against Vibrio vulnificus and Its Healing Potential on Infected Mice with Full-Thickness Excised Skin.

Shikonin has anticancer, anti-inflammatory, and wound healing activities. Vibrio vulnificus is an important marine foodborne pathogen with a high fatality rate and rapid pathogenesis that can infect humans through ingestion and wounds. In this study, the antibacterial activity and possible antibacterial mechanism of shikonin against V. vulnificus were investigated. In addition, the ability of shikonin to control V. vulnificus infection in both pathways was assessed by artificially contaminated oysters and full-thickness excised skin-infected mice. Shikonin treatment can cause abnormal cell membrane function, as evidenced by hyperpolarization of the cell membrane, significant decreased intracellular ATP concentration (p < 0.05), significant increased intracellular reactive oxygen species and malondialdehyde content (p < 0.05), decreased cell membrane integrity, and changes in cell morphology. Shikonin at 40 and 80 μg/mL reduced bacterial numbers in shikonin-contaminated oysters by 3.58 and 2.18 log colony-forming unit (CFU)/mL. Shikonin can promote wound healing in mice infected with V. vulnificus by promoting the formation of granulation tissue, hair follicles, and sebaceous glands, promoting epithelial cell regeneration and epidermal growth factor production. These findings suggest that shikonin has a strong inactivation effect on V. vulnificus and can be used in food production and wound healing to effectively control V. vulnificus and reduce the number of diseases associated with it.

Antibacterial Activity and Mechanism of Oxidized Bacterial Nanocellulose with Different Carboxyl Content.

Oxidized bacterial nanocellulose (OBC) is reported to prevent microbial growth, but its antibacterial characteristics and mechanism are still unclear. Here, the antibacterial mechanism of OBC is explored by detecting and assessing the interaction of OBC with different carboxyl content on Staphylococcus aureus and Escherichia coli. The results show that OBC has strong antibacterial activity and antibiofilm activity against S. aureus and E. coli, which is positively correlated with the carboxyl content of OBC. After OBC treatment, the bacteria adhesion is inhibited and the cell membrane is destroyed leading to increased permeability. Further investigation reveals that the concentration of cyclic diguanosine monophosphate (c-di-GMP)that induced biofilm formation is significantly decreased to 1.81pmolmg-1 after OBC treatment. In addition, OBC inactivates mature biofilms, with inactivation rates up to 79.3%. This study suggests that OBC has excellent antibacterial and antiadhesion properties, which can increase the cell membrane permeability and inhibit c-di-GMP formation. In addition, OBC also has a strong inactivation effect on mature biofilm, which can be used as an effective antibiofilm agent.

Peracetic acid can be used as a disinfectant for gilthead sea bream (Sparus aurata) juveniles without affecting fish welfare

Peracetic acid (PAA) is a European Union Commission authorized disinfectant for use in animal health care. It has shown a strong inactivation potential for bacteria, viruses, fungi and bacterial spores. A stress-related adaptive response after an exposure to PAA has been described in different species of fish such as carp (Cyprinus carpio), rainbow trout (Oncorhynchus mykiss). The present study aims to evaluate the response of gilthead seabream systemic (plasma) as mucosal (gill and skin) defences after PAA a unique exposure (5 min), by measuring cortisol, glucose and lactate in plasma, as well as the expression of several genes (glutathione peroxidase, glucocorticoid receptor, superoxide dismutase 2 and superoxide dismutase involved in the response to oxidative stress in mucosal tissues). study. We observe how seabream (Sparus aurata) copes with oxidative stress induced by PAA. PAA exposure did not induce an important antioxidant response in fish, whereas induced a mild response to stress, with a fast and effective recovery of basal levels after 24 h. Although PAA triggers a mild stress response, the response described in our study reflect that it can be used for sea bream in the concentration tested with no severe physiological consequences.

Cabernet Sauvignon Red Must Processing by UHPH to Produce Wine Without SO2: the Colloidal Structure, Microbial and Oxidation Control, Colour Protection and Sensory Quality of the Wine

A cryo-macerated must of V. vinifera L. cabernet sauvignon was processed by ultra-high-pressure homogenisation (UHPH) sterilisation without the use of SO2. The UHPH treatment of the must was carried out continuously at a pressure of 300 MPa and reaching a maximum temperature of 77 °C for less than 0.2 s. The colloidal structure of the UHPH must was evaluated by atomic force microscopy (AFM) measuring an average particle size of 457 nm. The initial microbial load was 4-log CFU/mL (yeast), 3-log CFU/mL (bacteria). No yeast and non-sporulating bacteria were detected in 1 mL and 10 mL of the UHPH-treated must, respectively. Furthermore, no fermentative activity was detected in the non-inoculated UHPH-treated musts for more than 50 days. A strong inactivation of the oxidative enzymes was observed, with lower oxidation (≈ × 3) than controls. The antioxidant activity of the UHPH-treated must was much higher (106%) than that of the control must. UHPH had a protective effect in total anthocyanins, and especially in acylated anthocyanins (+ 9.3%); furthermore, the fermentation produces fewer higher alcohol (-44,3%) and more 2-phenylethyl acetate (+ 63%).

Effect of ultrasonic treatment on the activity of sugar metabolism relative enzymes and quality of coconut water

In this study, tender coconuts were treated with high-intensity ultrasound (US) for 20 min at a frequency of 20 kHz and a power of 2400 W. Compared with control group, US treated coconut water had a higher content of total soluble solid and sugar/acid ratio along with a lower pH value and conductivity, and the contents of sucrose, fructose and glucose were also higher. Results from HS-SPME/GC–MS showed that there was no significant difference in the content of volatile compounds in coconut water before and after US treatment. The activities of sugar metabolism enzymes such as sucrose phosphate synthase, sucrose synthase, acid invertase (AI) and neutral invertase were inhibited by US, of which AI had the strongest inactivation. Circular dichroism and fluorescence spectra showed that the secondary and tertiary structure of AI molecule were destroyed with the increase of US intensity and time, which was confirmed by the change of particle size distribution pattern and scanning electron microscopy. Molecular docking and molecular dynamics showed that US treatment prevented the recognition and binding of sucrose and AI molecules, thereby inhibiting the decomposition of sucrose. In conclusion, our results indicate that US can inhibit the activity of AI and maintain the sugar content to increase the quality as well as extend the shelflife of coconut water, which will bring more commercial value.

Abstract 1271: In vitro and in vivo characterization of BI 1823911 - a novel KRASG12C selective small molecule inhibitor

Abstract KRASG12C mutations are predominantly found in non-small cell lung cancer (NSCLC, 13%), in colorectal cancer (CRC, 3%), and with a lower prevalence in pancreatic ductal adenocarcinoma (PDAC, 1%). The amino acid exchange at position 12 from glycine to cysteine renders RAS insensitive to GAP-catalyzed hydrolysis but not to intrinsic hydrolysis and consequently, KRASG12C is still dependent on GEF stimulation to achieve full activation. The active GTP-loaded form of KRASG12C is favored and leads to activation of downstream signaling and proliferation. A number of recent publications has shown that targeting this mutant form of KRAS, using several covalent KRASG12C inhibitors binding to the inactive GDP-KRASG12C form, leads to anti-proliferative effects and induction of apoptosis in KRASG12C mutant cancer cell lines, CDX and PDX models. Early clinical data for AMG 510 and MRTX849 revealed a response rate of 35-45% in NSCLC and of 7-17% in CRC patients. Here, we show that BI 1823911 has potent anti-proliferative activity in a panel of KRASG12C mutant cancer cell lines with higher or similar potency compared to these two most advanced compounds in clinical development. In a panel of KRASG12C NSCLC cell lines, treatment with BI 1823911 results in downregulation of MAPK pathway-responsive genes, such as DUSP6 and CCND1, and the extent of pathway modulation correlates with sensitivity. Likewise, we observe strong and sustained inactivation of the MAPK pathway at the protein level using p-ERK as a pharmacodynamic (PD) biomarker. A MIA PaCa-2 cell line-derived pancreatic cancer xenograft model was selected for extensive PK/PD/efficacy analyses in vivo. Briefly, BI 1823911 tested at 60 mg/kg showed similar anti-tumor activity compared to both competitor compounds dosed at clinically relevant exposures. Results of the ongoing in-depth PK/PD/efficacy analysis will be shared. Furthermore, BI 1823911 was tested with a daily oral dose of 60 mg/kg in a panel of NSCLC or CRC CDX or PDX mouse models and showed comparable efficacy to AMG 510 and MRTX849, respectively. Preclinical and clinical data suggest that monotherapy with a KRASG12C inhibitor will not be sufficient to achieve durable response. Combination therapy of a KRASG12C inhibitor may therefore lead to enhanced anti-tumor efficacy and may address adaptive resistance mechanisms. Therefore, we selected a panel of KRASG12C mutant cancer cell lines and tested a large set of compounds in combination with BI 1823911 to identify synergistic anti-proliferative activity. Among other MAPK and PI3K pathway inhibitors, a SOS1::KRAS inhibitor was confirmed as promising combination partner. We will show results from in vitro and in vivo combination studies in NSCLC and CRC tumor models that show deep and durable responses upon combination of BI 1823911 with SOS1::KRAS inhibitor BI 1701963 providing a strong rationale for clinical investigation of this combination. Citation Format: Fabio Savarese, Andreas Gollner, Dorothea Rudolph, Jesse Lipp, Johannes Popow, Marco H. Hofmann, Heribert Arnhof, Jörg Rinnenthal, Francesca Trapani, Michael Gmachl, Daniel Gerlach, Joachim Broeker, Peter Ettmayer, Andreas Mantoulidis, Jason Phan, Christian A. Smethurst, Matthias Treu, Alex G. Waterson, Hengyu Lu, Annette Machado, Joseph Daniele, Stephan W. Fesik, Christopher P. Vellano, Timothy P. Heffernan, Joseph R. Marszalek, Darryl B. McConnell, Mark Petronczki, Norbert Kraut, Irene C. Waizenegger. In vitro and in vivo characterization of BI 1823911 - a novel KRASG12C selective small molecule inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1271.

Neuronal VCP loss of function recapitulates FTLD-TDP pathology.

SUMMARYThe pathogenic mechanism by which dominant mutations in VCP cause multisystem proteinopathy (MSP), a rare neurodegenerative disease that presents as fronto-temporal lobar degeneration with TDP-43 inclusions (FTLD-TDP), remains unclear. To explore this, we inactivate VCP in murine postnatal forebrain neurons (VCP conditional knockout [cKO]). VCP cKO mice have cortical brain atrophy, neuronal loss, autophago-lysosomal dysfunction, and TDP-43 inclusions resembling FTLD-TDP pathology. Conditional expression of a single disease-associated mutation, VCP-R155C, in a VCP null background similarly recapitulates features of VCP inactivation and FTLD-TDP, suggesting that this MSP mutation is hypomorphic. Comparison of transcriptomic and proteomic datasets from genetically defined patients with FTLD-TDP reveal that progranulin deficiency and VCP insufficiency result in similar profiles. These data identify a loss of VCP-dependent functions as a mediator of FTLD-TDP and reveal an unexpected biochemical similarity with progranulin deficiency.