BACKGROUND: Reactive oxygen species, such as superoxide (O(2)(-)), are involved in the abnormal growth of various cell types. Angiotensin II (Ang II) is one of the most potent inducers of oxidative stress in the vasculature. The molecular events involved in Ang II-induced proliferation of vascular smooth muscle cells (VSMCs) are ... More
Patient-derived glioblastoma organoids as real-time avatars for assessing responses to clinical CAR-T cell therapy
AuthorsMeghan Logun, Xin Wang, Yusha Sun, Stephen J Bagley, Nannan Li, Arati Desai, Daniel Y Zhang, MacLean P Nasrallah, Emily Ling-Lin Pai, Bike Su Oner, Gabriela Plesa, Donald Siegel, Zev A Binder, Guo-Li Ming, Hongjun Song, Donald M O'Rourke
JournalCell Stem Cell
PubMed ID39657679
Patient-derived tumor organoids have been leveraged for disease modeling and preclinical studies but rarely applied in real time to aid with interpretation of patient treatment responses in clinics. We recently demonstrated early efficacy signals in a first-in-human, phase 1 study of dual-targeting chimeric antigen receptor (CAR)-T cells (EGFR-IL13Rα2 CAR-T cells) ... More
Effects of an indole derivative on cell proliferation, transfection, and alternative splicing in production of lentiviral vectors by transient co-transfection.
AuthorsMier NC,Roper DK
JournalPloS one
PubMed ID38833479
Lentiviral vectors derived from human immunodeficiency virus type I are widely used to deliver functional gene copies to mammalian cells for research and gene therapies. Post-transcriptional splicing of lentiviral vector transgene in transduced host and transfected producer cells presents barriers to widespread application of lentiviral vector-based therapies. The present study ... More
Mutation of the cytoplasmic domain of the integrin beta 3 subunit. Differential effects on cell spreading, recruitment to adhesion plaques, endocytosis, and phagocytosis.
Authors Ylänne J; Huuskonen J; O'Toole T E; Ginsberg M H; Virtanen I; Gahmberg C G;
JournalJ Biol Chem
PubMed ID7721884
'The cytoplasmic domain of the beta subunit of the alpha IIb beta 3 integrin is required for cell spreading on fibrinogen. Here we report that deletion of six amino acids from the COOH terminus of the beta 3 (I757TYRGT) totally abolished cell spreading and formation of adhesion plaques, whereas retaining ... More
Gene correction of the apolipoprotein (Apo) E2 phenotype to wild-type ApoE3 by in situ chimeraplasty.
AuthorsTagalakis AD, Graham IR, Riddell DR, Dickson JG, Owen JS,
JournalJ Biol Chem
PubMed ID11278248
'Apolipoprotein (apo) E is a polymorphic plasma protein, synthesized mainly by liver. Here, we evaluate whether synthetic DNA-RNA oligonucleotides (chimeraplasts) can convert a dysfunctional isoform, apoE2 (C --> T, R158C), which causes Type III hyperlipidemia and premature atherosclerosis, into apoE3. First, we treated recombinant Chinese hamster ovary cells stably secreting ... More
Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells.
AuthorsZhang J, Nuebel E, Wisidagama DR, Setoguchi K, Hong JS, Van Horn CM, Imam SS, Vergnes L, Malone CS, Koehler CM, Teitell MA,
JournalNat Protoc
PubMed ID22576106
Measurements of glycolysis and mitochondrial function are required to quantify energy metabolism in a wide variety of cellular contexts. In human pluripotent stem cells (hPSCs) and their differentiated progeny, this analysis can be challenging because of the unique cell properties, growth conditions and expense required to maintain these cell types. ... More
Reprogramming human fibroblasts to pluripotency using modified mRNA.
AuthorsMandal PK, Rossi DJ,
JournalNat Protoc
PubMed ID23429718
Induced pluripotent stem (iPS) cells hold the potential to revolutionize regenerative medicine through their capacity to generate cells of diverse lineages for future patient-specific cell-based therapies. To facilitate the transition of iPS cells to clinical practice, a variety of technologies have been developed for transgene-free pluripotency reprogramming. We recently reported ... More
Phosphorylation of human Rad9 is required for genotoxin-activated checkpoint signaling.
AuthorsRoos-Mattjus P, Hopkins KM, Oestreich AJ, Vroman BT, Johnson KL, Naylor S, Lieberman HB, Karnitz LM,
JournalJ Biol Chem
PubMed ID12709442
Rad9, a key component of genotoxin-activated checkpoint signaling pathways, associates with Hus1 and Rad1 in a heterotrimeric complex (the 9-1-1 complex). Rad9 is inducibly and constitutively phosphorylated. However, the role of Rad9 phosphorylation is unknown. Here we identified nine phosphorylation sites, all of which lie in the carboxyl-terminal 119-amino acid ... More
Efficient intracellular assembly of papillomaviral vectors.
Although the papillomavirus structural proteins, L1 and L2, can spontaneously coassemble to form virus-like particles, currently available methods for production of L1/L2 particles capable of transducing reporter plasmids into mammalian cells are technically demanding and relatively low-yield. In this report, we describe a simple 293 cell transfection method for efficient ... More
Hypoxia and Nitric Oxide Treatment Confer Tolerance to Glucose Starvation in a 5'-AMP-activated Protein Kinase-dependent Manner.
Hypoxia is a critical event for higher organisms, and cells and tissues react by increasing the oxygen supply by vasodilatation, angiogenesis, and erythropoiesis and maintaining cellular energy by increasing glycolysis and inhibiting anabolic pathways. Stimulation of glycolysis has been regarded as the main response that increases energy production during hypoxia; ... More
Purification, Cloning, and Characterization of Nek8, a Novel NIMA-related Kinase, and Its Candidate Substrate Bicd2.
Authors Holland Pamela M; Milne Alison; Garka Kirsten; Johnson Richard S; Willis Cynthia; Sims John E; Rauch Charles T; Bird Timothy A; Virca G Duke;
JournalJ Biol Chem
PubMed ID11864968
We describe the isolation, cloning, and characterization of human Nek8, a new mammalian NIMA-related kinase, and its candidate substrate Bicd2. Nek8 was isolated as a beta-casein kinase activity in rabbit lung and has an N-terminal catalytic domain homologous to the Nek family of protein kinases. Nek8 also contains a central ... More
Human Synovial MSC Good Manufacturing Practices for Articular Cartilage Regeneration.
AuthorsFernandes TL, Kimura HA, Pinheiro CCG, Shimomura K, Nakamura N, Ferreira JRM, Gomoll AH, Hernandez AJ, Bueno DF,
JournalTissue Eng Part C Methods
PubMed ID30412046
cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions. Chondral lesion is a pathology with high prevalence, reaching as much as 63% of general population and 36% among athletes. Despite Autologous Chondrocyte Implantation (ACI) versatility, it still fails to fully reproduce hyaline articular cartilage characteristics. Mesenchymal ... More