Dendrimer-Based Selective Proteostasis-Inhibition Strategy to Control NSCLC Growth and Progression.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Gene Expression Regulation, Neoplastic*; Adenosine Triphosphatases/*antagonists & inhibitors ; Antineoplastic Agents/*pharmacology ; Apoptosis/*drug effects ; Cell Cycle Proteins/*antagonists & inhibitors ; Enzyme Inhibitors/*pharmacology ; Epithelial Cells/*drug effects ; Quinazolines/*pharmacology; Adenosine Triphosphatases/genetics ; Adenosine Triphosphatases/metabolism ; Antineoplastic Agents/chemistry ; Bronchi/drug effects ; Bronchi/metabolism ; Bronchi/pathology ; Caspase 3/genetics ; Caspase 3/metabolism ; Caspase 7/genetics ; Caspase 7/metabolism ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Cell Line, Tumor ; Cell Movement/drug effects ; Cell Proliferation/drug effects ; Dendrimers/chemistry ; Dendrimers/metabolism ; Drug Carriers ; Drug Compounding ; Enzyme Inhibitors/chemistry ; Epithelial Cells/metabolism ; Epithelial Cells/pathology ; G2 Phase Cell Cycle Checkpoints/drug effects ; Humans ; Proteolysis/drug effects ; Quinazolines/chemistry ; Receptors, Peptide/genetics ; Receptors, Peptide/metabolism ; Ubiquitinated Proteins/genetics ; Ubiquitinated Proteins/metabolism ; Valosin Containing Protein

Elevated valosin containing protein (VCP/p97) levels promote the progression of non-small cell lung carcinoma (NSCLC). Although many VCP inhibitors are available, most of these therapeutic compounds have low specificity for targeted tumor cell delivery. Hence, the primary aim of this study was to evaluate the in vitro efficacy of dendrimer-encapsulated potent VCP-inhibitor drug in controlling non-small cell lung carcinoma (NSCLC) progression. The VCP inhibitor(s) (either in their pure form or encapsulated in generation-4 PAMAM-dendrimer with hydroxyl surface) were tested for their in vitro efficacy in modulating H1299 (NSCLC cells) proliferation, migration, invasion, apoptosis and cell cycle progression. Our results show that VCP inhibition by DBeQ was significantly more potent than NMS-873 as evident by decreased cell proliferation (p<0.0001, MTT-assay) and migration (p<0.05; scratch-assay), and increased apoptosis (p<0.05; caspase-3/7-assay) as compared to untreated control cells. Next, we found that dendrimer-encapsulated DBeQ (DDNDBeQ) treatment increased ubiquitinated-protein accumulation in soluble protein-fraction (immunoblotting) of H1299 cells as compared to DDN-control, implying the effectiveness of DBeQ in proteostasis-inhibition. We verified by immunostaining that DDNDBeQ treatment increases accumulation of ubiquitinated-proteins that co-localizes with an ER-marker, KDEL. We observed that proteostasis-inhibition with DDNDBeQ, significantly decreased cell migration rate (scratch-assay and transwell-invasion) as compared to the control-DDN treatment (p<0.05). Moreover, DDNDBeQ treatment showed a significant decrease in cell proliferation (p<0.01, MTT-assay) and increased caspase-3/7 mediated apoptotic cell death (p<0.05) as compared to DDN-control. This was further verified by cell cycle analysis (propidium-iodide-staining) that demonstrated significant cell cycle arrest in the G2/M-phase (p<0.001) by DDNDBeQ treatment as compared to control-DDN. Moreover, we confirmed by clonogenic-assay that DDNDBeQ treatment significantly (p<0.001) inhibits H1299 colony-formation as compared to control/DDN. Overall, encapsulation of potent VCP-inhibitor DBeQ into a dendrimer allows selective VCP-mediated proteostasis-inhibition for controlling NSCLC-tumor growth and progression to allow tumor-targeted sustained drug delivery.

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R43 CA141868 United States CA NCI NIH HHS

0 (Antineoplastic Agents)
0 (Cell Cycle Proteins)
0 (Dendrimers)
0 (Drug Carriers)
0 (Enzyme Inhibitors)
0 (KDELR1 protein, human)
0 (N2,N4-dibenzylquinazoline-2,4-diamine)
0 (PAMAM Starburst)
0 (Quinazolines)
0 (Receptors, Peptide)
0 (Ubiquitinated Proteins)
EC 3.4.22.- (CASP3 protein, human)
EC 3.4.22.- (CASP7 protein, human)
EC 3.4.22.- (Caspase 3)
EC 3.4.22.- (Caspase 7)
EC 3.6.1.- (Adenosine Triphosphatases)
EC 3.6.4.6 (VCP protein, human)
EC 3.6.4.6 (Valosin Containing Protein)

Date Created: 20160720 Date Completed: 20170718 Latest Revision: 20201215

20240829

PMC4951140

10.1371/journal.pone.0158507

27434122