Genetic and drug screens applied to a Drosophila model of acute myeloid leukemia reveal potential link to hypoxia signaling

Dis Model Mech. 2021 Sep 28:dmm.048953. doi: 10.1242/dmm.048953.  

Pharmacological or genetic inhibition of hypoxia signaling attenuates oncogenic
RAS-induced cancer phenotypes.

Zhu JY, Huang X, Fu Y, Wang Y, Zheng P, Liu Y, Han Z

Author information:
(1)Center for Precision Disease Modeling, Department of Medicine, University of
Maryland School of Medicine, Baltimore, MD 21201, USA.
(2)Division of Endocrinology, Diabetes and Nutrition, Department of Medicine,
University of Maryland School of Medicine, Baltimore, MD 21201, USA.
(3)Division of Immunotherapy, University of Maryland School of Medicine,
Baltimore, MD 21201, USA.

Abstract: "Oncogenic Ras mutations are highly prevalent in hematopoietic malignancies. However, it is difficult to directly target oncogenic RAS proteins for therapeutic intervention. We have developed a Drosophila Acute Myeloid Leukemia (AML) model induced by human KRASG12V, which exhibits a dramatic increase in myeloid-like leukemia cells. We performed both genetic and drug screens using this model. The genetic screen identified 24 candidate genes able to attenuate the oncogenic RAS-induced phenotype, including two key hypoxia pathway genes HIF1A and ARNT (HIF1B). The drug screen revealed echinomycin, an inhibitor of HIF1A, could effectively attenuate the leukemia phenotype caused by KRASG12V. Furthermore, we showed that echinomycin treatment could effectively suppress oncogenic RAS-driven leukemia cell proliferation using both human leukemia cell lines and a mouse xenograft model. These data suggest that inhibiting the hypoxia pathway could be an effective treatment approach for oncogenic RAS-induced cancer phenotype, and that echinomycin is a promising targeted drug to attenuate oncogenic RAS-induced cancer phenotypes."

DOI: 10.1242/dmm.048953
PMID: 34580712

Identification of protein interactors with fly GOLPH3 points to potential targets for cancer and other therapeutics

Cells. 2021 Sep 6;10(9):2336. doi: 10.3390/cells10092336.

Identification of GOLPH3 Partners in Drosophila Unveils Potential Novel Roles in Tumorigenesis and Neural Disorders.

Sechi S, Karimpour-Ghahnavieh A, Frappaolo A, Di Francesco L, Piergentili R, Schininà E, D'Avino PP, Giansanti MG

Author information:
(1)Istituto di Biologia e Patologia Molecolari del CNR, c/o Dipartimento di
Biologia e Biotecnologie, Sapienza Università di Roma, Piazzale A. Moro 5, 00185
Roma, Italy.
(2)Dipartimento di Scienze Biochimiche A. Rossi Fanelli, Sapienza Università di
Roma, Piazzale A. Moro 5, 00185 Roma, Italy.
(3)Department of Pathology, University of Cambridge, Tennis Court Road,
Cambridge CB2 1QP, UK.

Abstract: "Golgi phosphoprotein 3 (GOLPH3) is a highly conserved peripheral membrane protein localized to the Golgi apparatus and the cytosol. GOLPH3 binding to Golgi membranes depends on phosphatidylinositol 4-phosphate [PI(4)P] and regulates Golgi architecture and vesicle trafficking. GOLPH3 overexpression has been correlated with poor prognosis in several cancers, but the molecular mechanisms that link GOLPH3 to malignant transformation are poorly understood. We recently showed that PI(4)P-GOLPH3 couples membrane trafficking with contractile ring assembly during cytokinesis in dividing Drosophila spermatocytes. Here, we use affinity purification coupled with mass spectrometry (AP-MS) to identify the protein-protein interaction network (interactome) of Drosophila GOLPH3 in testes. Analysis of the GOLPH3 interactome revealed enrichment for proteins involved in vesicle-mediated trafficking, cell proliferation and cytoskeleton dynamics. In particular, we found that dGOLPH3 interacts with the Drosophila orthologs of Fragile X mental retardation protein and Ataxin-2, suggesting a potential role in the pathophysiology of disorders of the nervous system. Our findings suggest novel molecular targets associated with GOLPH3 that might be relevant for therapeutic intervention in cancers and other human diseases."

DOI: 10.3390/cells10092336
PMID: 34571985

Review: Drosophila accessory gland as a model organ for studies related to prostate cancer

Cells. 2021 Sep 10;10(9):2387. doi: 10.3390/cells10092387.

Drosophila Accessory Gland: A Complementary In Vivo Model to Bring New Insight  to Prostate Cancer.

Rambur A, Vialat M, Beaudoin C, Lours-Calet C, Lobaccaro JM, Baron S, Morel L, de Joussineau C

Author information:
(1)Rosalind and Morris Goodman Cancer Research Centre, McGill University, 1160 Pine Avenue West, Montréal, QC H3A 1A3, Canada.
(2)Institut de Génétique, Reproduction et Développement, Université Clermont Auvergne, CNRS UMR6293, INSERM U1103, 28 Place Henri Dunant, BP38, 63001 Clermont-Ferrand, France.

Abstract: "Prostate cancer is the most common cancer in aging men. ... there are still few effective treatments to cure its aggressive and metastatic stages. ... multiple research models, such as cell culture and mouse models, have been developed over the years and have improved our comprehension of the biology of the disease. Recently, a new model has been added with the use of the Drosophila accessory gland. ... this functional equivalent of the prostate represents a powerful, inexpensive, and rapid in vivo model to study epithelial carcinogenesis. The purpose of this review is to quickly overview the existing prostate cancer models, including their strengths and limitations. In particular, we discuss how the Drosophila accessory gland can be integrated as a convenient complementary model ..."

DOI: 10.3390/cells10092387
PMID: 34572036

Fly disease model of Limb-girdle muscular dystrophy D2 (LGMDDs)

Blázquez-Bernal Á, Fernandez-Costa JM, Bargiela A, Artero R. Inhibition of autophagy rescues muscle atrophy in a LGMDD2 Drosophila model. FASEB J. 2021 Oct;35(10):e21914. doi: 10.1096/fj.202100539RR. PMID: 34547132.

From the abstract: "Limb-girdle muscular dystrophy D2 (LGMDD2) is an ultrarare autosomal dominant myopathy caused by mutation of ... TNPO3 ... . ... Here we report the first animal model of the disease by expressing the human mutant TNPO3 gene in Drosophila musculature or motor neurons and concomitantly silencing the endogenous expression of the fly protein ortholog. A similar genotype expressing wildtype TNPO3 served as a control. Phenotypes characterization revealed that mutant TNPO3 expression targeted at muscles or motor neurons caused LGMDD2-like phenotypes such as muscle degeneration and atrophy, and reduced locomotor ability. ... Upregulated muscle autophagy observed in LGMDD2 patients was also confirmed in the fly model, in which the anti-autophagic drug chloroquine was able to rescue histologic and functional phenotypes. Overall, we provide a proof of concept of autophagy as a target to treat disease phenotypes and propose a neurogenic component to explain mutant TNPO3 pathogenicity in diseased muscles."

Multi-species study looks at potential treatment of Anthrax

ACS Infect Dis. 2021 Aug 13;7(8):2176-2191. doi: 10.1021/acsinfecdis.1c00190.

In Vivo Activity of Repurposed Amodiaquine as a Host-Targeting Therapy for the Treatment of Anthrax.

Martchenko Shilman M, Bartolo G, Alameh S, Peterson JW, Lawrence WS, Peel JE, Sivasubramani SK, Beasley DWC, Cote CK, Demons ST, Halasahoris SA, Miller LL, Klimko CP, Shoe JL, Fetterer
DP, McComb R, Ho CC, Bradley KA, Hartmann S, Cheng LW, Chugunova M, Kao CY, Tran JK, Derbedrossian A, Zilbermintz L, Amali-Adekwu E, Levitin A, West J

Anthrax is caused by Bacillus anthracis and can result in nearly 100% mortality due in part to anthrax toxin. Antimalarial amodiaquine (AQ) acts as a host-oriented inhibitor of anthrax toxin endocytosis. Here, we determined the pharmacokinetics and safety of AQ in mice, rabbits, and humans as well as the efficacy in the fly, mouse, and rabbit models of anthrax infection. In the therapeutic-intervention studies, AQ nearly doubled the survival of mice infected subcutaneously with a B. anthracis dose lethal to 60% of the animals (LD60). In rabbits challenged with 200 LD50 of aerosolized B. anthracis, AQ as a monotherapy delayed death, doubled the survival rate of infected animals that received a suboptimal amount of antibacterial levofloxacin, and reduced bacteremia and toxemia in tissues. Surprisingly, the anthrax efficacy of AQ relies on an additional host macrophage-directed antibacterial mechanism, which was validated in the toxin-independent Drosophila model of Bacillus infection. Lastly, a systematic literature review of the safety and pharmacokinetics of AQ in humans from over 2 000 published articles revealed that AQ is likely safe when taken as prescribed, and its pharmacokinetics predicts anthrax efficacy in humans. Our results support the future examination of AQ as adjunctive therapy for the prophylactic anthrax treatment.

DOI: 10.1021/acsinfecdis.1c00190
PMCID: PMC8369491
PMID: 34218660 [Indexed for MEDLINE]

Conflict of interest statement: The authors declare the following competing financial interest(s): M.M.S. and L.Z. are co-inventors on the AQ patent for anthrax.

Multi-species study identifies inhibitory compound with potential use for Down Syndrome

Phytomedicine. 2021 Aug 1;92:153695. doi: 10.1016/j.phymed.2021.153695. Online
ahead of print.

Aristolactam BIII, a naturally derived DYRK1A inhibitor, rescues Down syndrome-related phenotypes.

Choi M, Kim AK, Ham Y, Lee JY, Kim D, Yang A, Jo MJ, Yoon E, Heo JN, Han SB, Ki MH, Lee KS, Cho S

BACKGROUND: Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) is a significant pathogenic factor in Down syndrome (DS), wherein DYRK1A is overexpressed by 1.5-fold because of trisomy of human chromosome 21. Thus, DYRK1A inhibition is considered a therapeutic strategy to modify the disease.
PURPOSE: This study aims to identify a novel DYRK1A inhibitor and validate its therapeutic potential in DS-related pathological conditions.
 

STUDY DESIGN: In order to identify a novel DYRK1A inhibitor, we carried out two-step screening: a structure-based virtual screening of > 300,000 chemical library (first step) and cell-based nuclear factor of activated T-cells (NFAT)-response element (RE) promoter assay (second step). Primary hits were evaluated for their DYRK1A inhibitory activity using in vitro kinase assay and Tau phosphorylation in mammalian cells. Confirmed hit was further evaluated in pathological conditions including DYRK1A-overexpressing fibroblasts, flies, and mice.
 

RESULTS: We identified aristolactam BIII, a natural product derived from herbal plants, as a novel DYRK1A inhibitor. It potently inhibited the kinase activity of DYRK1A in vitro (IC50 = 9.67 nM) and effectively suppressed DYRK1A-mediated hyperphosphorylation of Tau in mammalian cells. Aristolactam BIII rescued the proliferative defects of DYRK1A transgenic (TG) mouse-derived fibroblasts and neurological and phenotypic defects of DS-like Drosophila models. Oral administration of aristolactam BIII acutely suppressed Tau hyperphosphorylation in the brain of DYRK1A TG mice. In the open field test, aristolactam BIII significantly ameliorated the exploratory behavioral deficit of DYRK1A TG mice.
 

CONCLUSION: Our work revealed that aristolactam BIII as a novel DYRK1A inhibitor rescues DS phenotypes in cells and in vivo and suggested its therapeutic potential for the treatment of DYRK1A-related diseases.

DOI: 10.1016/j.phymed.2021.153695
PMID: 34500300