Similarities and distinctions in the activation of the Pdr1 regulatory pathway by azole and non-azole drugs.

Unlabelled: Incidences of fluconazole (FLC) resistance among clinical isolates are a growing issue in clinics. The pleiotropic drug response network in confers azole resistance and is defined primarily by the ZnCys zinc cluster-containing transcription factor Pdr1 and target genes such as , which encodes an ATP-binding cassette transporter protein thought to act as an FLC efflux pump. Mutations in the gene that render the transcription factor hyperactive are the most common cause of fluconazole resistance among clinical isolates.

Similarities and distinctions in the activation of the Pdr1 regulatory pathway by azole and non-azole drugs.

Incidences of fluconazole (FLC) resistance among clinical isolates is a growing issue in clinics. The pleiotropic drug response (PDR) network in . confers azole resistance and is defined primarily by the ZnCys zinc cluster-containing transcription factor Pdr1 and target genes such as , that encodes an ATP-binding cassette transporter protein thought to act as a FLC efflux pump.

A series of pyrimidine-based antifungals with anti-mold activity disrupt ER function in .

Fungal infections are a major contributor to morbidity and mortality among immunocompromised populations. Moreover, fungal disease caused by molds are difficult to treat and are associated with particularly high mortality. To address the need for new mold-active antifungal drugs, we performed a high-throughput screen with , the most common pathogenic mold.

Development of a murine model to study the cerebral pathogenesis of .

Molds are environmental fungi that can cause disease in immunocompromised individuals. The most common pathogenic mold is which is typically inhaled into the lungs and causes invasive pulmonary disease. In a subset of these patients, this infection can spread from the lungs to other organs including the brain, resulting in cerebral aspergillosis.

The anti-cancer efficacy of a novel phenothiazine derivative is independent of dopamine and serotonin receptor inhibition.

Introduction: An attractive, yet unrealized, goal in cancer therapy is repurposing psychiatric drugs that can readily penetrate the blood-brain barrier for the treatment of primary brain tumors and brain metastases. Phenothiazines (PTZs) have demonstrated anti-cancer properties through a variety of mechanisms. However, it remains unclear whether these effects are entirely separate from their activity as dopamine and serotonin receptor (DR/5-HTR) antagonists.

CWHM-974 is a fluphenazine derivative with improved antifungal activity against due to reduced susceptibility to multidrug transporter-mediated resistance mechanisms.

Multidrug resistance (MDR) transporters such as ATP-Binding Cassette (ABC) and Major Facilitator Superfamily proteins are important mediators of antifungal drug resistance, particularly with respect to azole class drugs. Consequently, identifying molecules that are not susceptible to this mechanism of resistance is an important goal for new antifungal drug discovery. As part of a project to optimize the antifungal activity of clinically used phenothiazines, we synthesized a fluphenazine derivative (CWHM-974) with 8-fold higher activity against spp.

CWHM-974 is a fluphenazine derivative with improved antifungal activity against due to reduced susceptibility to multidrug transporter-mediated resistance mechanisms.

Multidrug resistance (MDR) transporters such as ATP Binding Cassette (ABC) and Major Facilitator Superfamily (MFS) proteins are important mediators of antifungal drug resistance, particularly with respect to azole class drugs. Consequently, identifying molecules that are not susceptible to this mechanism of resistance is an important goal for new antifungal drug discovery. As part of a project to optimize the antifungal activity of clinically used phenothiazines, we synthesized a fluphenazine derivative (CWHM-974) with 8-fold higher activity against spp.

Novel Keto-Alkyl-Pyridinium Antifungal Molecules Active in Models of Candida albicans Vascular Catheter Infection and Candida auris Skin Colonization.

New antifungal therapies are needed for both systemic, invasive infections in addition to superficial infections of mucosal and skin surfaces as well as biofilms associated with medical devices. The resistance of biofilm and biofilm-like growth phases of fungi contributes to the poor efficacy of systemic therapies to nonsystemic infections. Here, we describe the identification and characterization of a novel keto-alkyl-pyridinium scaffold with broad spectrum activity (2 to 16 μg/mL) against medically important yeasts and molds, including clinical isolates resistant to azoles and/or echinocandins.

A Dual-Readout High-Throughput Screening Assay for Small Molecules Active Against Aspergillus Fumigatus.

Human fungal infections caused by molds have been on the rise in recent years. These infections have high mortality rates compared to other fungal infections, and yet treatment options are limited due to resistance to clinical antifungals and lack of broad-spectrum activity against molds. Technical challenges associated with molds have limited large-throughput screening efforts for anti-mold compounds: therefore, we adapted an assay for use with A.

Novel keto-alkyl-pyridinium antifungal molecules active in models of in vivo vascular catheter infection and ex vivo skin colonization.

New antifungal therapies are needed for both systemic, invasive infections as well as superficial infections of mucosal and skin surfaces as well as biofilms associated with medical devices. The resistance of biofilm and biofilm-like growth phases of fungi contributes to the poor efficacy of systemic therapies to non-systemic infections. Here, we describe the identification and characterization of a novel keto-alkyl-pyridinium scaffold with broad spectrum activity (2-16 µg/mL) against medically important yeasts and moulds, including clinical isolates resistant to azoles and/or echinocandins.

Genetic Interaction Analysis Reveals that Cryptococcus neoformans Utilizes Multiple Acetyl-CoA-Generating Pathways during Infection.

Cryptococcus neoformans is an important human fungal pathogen for which the external environment is its primary niche. Previous work has shown that two nonessential acetyl-CoA metabolism enzymes, ATP-citrate lyase () and acetyl-CoA synthetase (), play important roles in C. neoformans infection.

Is Required for Cryptococcus neoformans Fitness : Application of Copper-Regulated Gene Expression to Mouse Models of Cryptococcosis.

There is an urgent need for new antifungals to treat cryptococcal meningoencephalitis, a leading cause of mortality in people living with HIV/AIDS. An important aspect of antifungal drug development is the validation of targets to determine whether they are required for the survival of the organism in animal models of disease. In Cryptococcus neoformans, a copper-regulated promoter (pCTR4-2) has been used previously to modulate gene expression .

Gluconate Kinase Is Required for Gluconate Assimilation and Sporulation in Cryptococcus neoformans.

Cryptococcus neoformans is an environmental yeast and an opportunistic human pathogen. The ability to cause disease depends on the ability to adapt to the human host. Previous studies implicated nfectivity-elated inase (, CNAG_03048) as required for establishing an infection.

An Alanine Aminotransferase Is Required for Biofilm-Specific Resistance of Aspergillus fumigatus to Echinocandin Treatment.

Alanine metabolism has been suggested as an adaptation strategy to oxygen limitation in organisms ranging from plants to mammals. Within the pulmonary infection microenvironment, Aspergillus fumigatus forms biofilms with steep oxygen gradients defined by regions of oxygen limitation. An alanine aminotransferase, AlaA, was observed to function in alanine catabolism and is required for several aspects of A.

Genetic analysis of Hsp90 function in Cryptococcus neoformans highlights key roles in stress tolerance and virulence.

The opportunistic human fungal pathogen Cryptococcus neoformans has tremendous impact on global health, causing 181,000 deaths annually. Current treatment options are limited, and the frequent development of drug resistance exacerbates the challenge of managing invasive cryptococcal infections. In diverse fungal pathogens, the essential molecular chaperone Hsp90 governs fungal survival, drug resistance, and virulence.

A Unique Dual-Readout High-Throughput Screening Assay To Identify Antifungal Compounds with Aspergillus fumigatus.

Treatment of invasive mold infections is limited by the lack of adequate drug options that are effective against these fatal infections. High-throughput screening of molds using traditional antifungal assays of growth is problematic and has greatly limited our ability to identify new mold-active agents. Here, we present a high-throughput screening platform for use with Aspergillus fumigatus, the most common causative agent of invasive mold infections, for the discovery of novel mold-active antifungals.

Enantioselective Synthesis of (+)-Hippolide J and Reevaluation of Antifungal Activity.

A synthesis of the reported antifungal agent (+)-hippolide J is presented. The rapid assembly of the natural product was enabled through implementation of an enantioselective isomerization/[2 + 2]-cycloaddition sequence. Due to the simplicity of the route, >100 mg of the natural product were prepared in a single pass.

Antifungal drug screening: thinking outside the box to identify novel antifungal scaffolds.

Invasive fungal infections are responsible for a significant disease burden worldwide. Drugs to treat these infections are limited to only four unique classes, and despite these available treatments, mortality rates remain unacceptably high. In this review, we will discuss antifungal drug screening and how the approach to identifying novel compounds needs move away from traditional growth-based assays in order to meet the demand for new drugs.

Benzothiourea Derivatives Target the Secretory Pathway of the Human Fungal Pathogen .

is one of the most important human fungal pathogens and causes life-threatening meningoencephalitis in immunocompromised patients. The current gold standard therapy for meningoencephalitis is based on medications that are over 50 years old and is not readily available in regions with high disease burden. Here, we report the mycologic, mechanistic, and pharmacologic characterization of a set of benzothioureas with highly selective fungicidal activity against .

Host Carbon Dioxide Concentration Is an Independent Stress for Cryptococcus neoformans That Affects Virulence and Antifungal Susceptibility.

The ability of to cause disease in humans varies significantly among strains with highly related genotypes. In general, environmental isolates of pathogenic species such as var. have reduced virulence relative to clinical isolates, despite having no differences in the expression of the canonical virulence traits (high-temperature growth, melanization, and capsule formation).

Characterizing the Pathogenic, Genomic, and Chemical Traits of , a Close Relative of the Major Human Fungal Pathogen .

is closely related to , the major cause of invasive mold infections. Even though is commonly found in diverse environments, including hospitals, it rarely causes invasive disease. Why causes less human disease than is unclear.

Response to Comment on "Sterilizing immunity in the lung relies on targeting fungal apoptosis-like programmed cell death".

Aouacheria question the interpretation of contemporary assays to monitor programmed cell death with apoptosis-like features (A-PCD) in Although our study focuses on fungal A-PCD for host immune surveillance and infectious outcomes, the experimental approach incorporates multiple independent A-PCD markers and genetic manipulations based on fungal rather than mammalian orthologs to circumvent the limitations associated with any single approach.

Interleukin 1α Is Critical for Resistance against Highly Virulent Aspergillus fumigatus Isolates.

Heterogeneity among isolates results in unique virulence potential and inflammatory responses. How these isolates drive specific immune responses and how this affects fungally induced lung damage and disease outcome are unresolved. We demonstrate that the highly virulent CEA10 strain is able to rapidly germinate within the immunocompetent lung environment, inducing greater lung damage, vascular leakage, and interleukin 1α (IL-1α) release than the low-virulence Af293 strain, which germinates with a lower frequency in this environment.

Sterilizing immunity in the lung relies on targeting fungal apoptosis-like programmed cell death.

Humans inhale mold conidia daily and typically experience lifelong asymptomatic clearance. Conidial germination into tissue-invasive hyphae can occur in individuals with defects in myeloid function, although the mechanism of myeloid cell-mediated immune surveillance remains unclear. By monitoring fungal physiology in vivo, we demonstrate that lung neutrophils trigger programmed cell death with apoptosis-like features in conidia, the most prevalent human mold pathogen.