title: AshwaMAX and Withaferin A inhibits gliomas in cellular and murine orthotopic models.
abstract: Glioblastoma multiforme (GBM) is an aggressive, malignant cancer Johnson and O'Neill (J Neurooncol 107: 359-364, 2012). An extract from the winter cherry plant (Withania somnifera ), AshwaMAX, is concentrated (4.3 %) for Withaferin A; a steroidal lactone that inhibits cancer cells Vanden Berghe et al. (Cancer Epidemiol Biomark Prev 23: 1985-1996, 2014). We hypothesized that AshwaMAX could treat GBM and that bioluminescence imaging (BLI) could track oral therapy in orthotopic murine models of glioblastoma. Human parietal-cortical glioblastoma cells (GBM2, GBM39) were isolated from primary tumors while U87-MG was obtained commercially. GBM2 was transduced with lentiviral vectors that express Green Fluorescent Protein (GFP)/firefly luciferase fusion proteins. Mutational, expression and proliferative status of GBMs were studied. Intracranial xenografts of glioblastomas were grown in the right frontal regions of female, nude mice (n = 3-5 per experiment). Tumor growth was followed through BLI. Neurosphere cultures (U87-MG, GBM2 and GBM39) were inhibited by AshwaMAX at IC50 of 1.4, 0.19 and 0.22 µM equivalent respectively and by Withaferin A with IC50 of 0.31, 0.28 and 0.25 µM respectively. Oral gavage, every other day, of AshwaMAX (40 mg/kg per day) significantly reduced bioluminescence signal (n = 3 mice, p < 0.02, four parameter non-linear regression analysis) in preclinical models. After 30 days of treatment, bioluminescent signal increased suggesting onset of resistance. BLI signal for control, vehicle-treated mice increased and then plateaued. Bioluminescent imaging revealed diffuse growth of GBM2 xenografts. With AshwaMAX, GBM neurospheres collapsed at nanomolar concentrations. Oral treatment studies on murine models confirmed that AshwaMAX is effective against orthotopic GBM. AshwaMAX is thus a promising candidate for future clinical translation in patients with GBM.
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA. echangcv@stanford.edu.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA. echangcv@stanford.edu.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA. echangcv@stanford.edu.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA. echangcv@stanford.edu.
authors.forename: Edwin
authors.initials: E
authors.lastname: Chang
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Christoph
authors.initials: C
authors.lastname: Pohling
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Arutselvan
authors.initials: A
authors.lastname: Natarajan
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Timothy H
authors.initials: TH
authors.lastname: Witney
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Jasdeep
authors.initials: J
authors.lastname: Kaur
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Lingyun
authors.initials: L
authors.lastname: Xu
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Gayatri
authors.initials: G
authors.lastname: Gowrishankar
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Aloma L
authors.initials: AL
authors.lastname: D'Souza
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Surya
authors.initials: S
authors.lastname: Murty
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Sophie
authors.initials: S
authors.lastname: Schick
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Liyin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Wu
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Phoo
authors.initials: P
authors.lastname: Khaw
authors.affiliation: Department of Medicine, Ludwig Institute for Cancer Research, University of California at San Diego, La Jolla, California, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Mischel
authors.affiliation: Cellworks Group Inc., San Jose, CA, USA.
authors.forename: Taher
authors.initials: T
authors.lastname: Abbasi
authors.affiliation: Cellworks Group Inc., San Jose, CA, USA.
authors.forename: Shahabuddin
authors.initials: S
authors.lastname: Usmani
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA.
authors.forename: Parag
authors.initials: P
authors.lastname: Mallick
authors.affiliation: Departments of Radiology and Bioengineering, Stanford University, Palo Alto, CA, USA. sgambhir@stanford.edu.|Molecular Imaging Program at Stanford, Stanford University, Palo Alto, CA, USA. sgambhir@stanford.edu.|Canary Center at Stanford for Cancer Early Detection, Stanford University, Palo Alto, CA, USA. sgambhir@stanford.edu.|Nuclear Medicine, Stanford University, Palo Alto, CA, USA. sgambhir@stanford.edu.
authors.forename: Sanjiv S
authors.initials: SS
authors.lastname: Gambhir
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000494:pharmacology; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D066246:ErbB Receptors / Q000378:metabolism; D005260:Female; D005909:Glioblastoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008163:Luminescent Measurements; D051379:Mice; D008819:Mice, Nude; D058953:Neural Stem Cells / Q000187:drug effects; D010936:Plant Extracts / Q000008:administration & dosage* / Q000737:chemistry; D032341:Withania / Q000737:chemistry*; D054358:Withanolides / Q000008:administration & dosage* / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts; D054358:Withanolides; C512478:EGFR protein, human; D066246:ErbB Receptors; C009684:withaferin A
keywords: AshwaMAX; Bioluminescent; Glioblastoma; Preclinical; Withaferin A