GZ17-6.02 and Pemetrexed Interact to Kill Osimertinib-Resistant NSCLC Cells That Express Mutant ERBB1 Proteins

Abstract

We determined the molecular mechanisms by which the novel therapeutic GZ17-6.02 killed non-small cell lung cancer (NSCLC) cells. Erlotinib, afatinib, and osimertinib interacted with GZ17-6.02 to kill NSCLC cells expressing mutant EGFR proteins. GZ17-6.02 did not interact with any EGFR inhibitor to kill osimertinib-resistant cells. GZ17-6.02 interacted with the thymidylate synthase inhibitor pemetrexed to kill NSCLC cells expressing mutant ERBB1 proteins or mutant RAS proteins or cells that were resistant to EGFR inhibitors. The drugs interacted to activate ATM, the AMPK, and ULK1 and inactivate mTORC1, mTORC2, ERK1/2, AKT, eIF2α; and c-SRC. Knockdown of ATM or AMPKα1 prevented ULK1 activation. The drugs interacted to cause autophagosome formation followed by flux, which was significantly reduced by knockdown of ATM, AMPKα1, and eIF2α, or by expression of an activated mTOR protein. Knockdown of Beclin1, ATG5, or [BAX + BAK] partially though significantly reduced drug combination lethality as did expression of activated mTOR/AKT/MEK1 or over-expression of BCL-XL. Expression of dominant negative caspase 9 weakly reduced killing. The drug combination reduced the expression of HDAC2 and HDAC3, which correlated with lower PD-L1, IDO1, and ODC levels and increased MHCA expression. Collectively, our data support consideration of combining GZ17-6.02 and pemetrexed in osimertinib-resistant NSCLC.

GZ17-6.02 kills prostate cancer cells in vitro and in vivo

“…Olaparib was purchased from Selleckchem (Houston, TX). All Materials were obtained as described in the references (1)(2)(3)(4)(5)(6). Trypsin-EDTA, DMEM, RPMI, penicillin-streptomycin were purchased from GIBCOBRL (GIBCOBRL Life Technologies, Grand Island, NY).…”

“…Trypsin-EDTA, DMEM, RPMI, penicillin-streptomycin were purchased from GIBCOBRL (GIBCOBRL Life Technologies, Grand Island, NY). Other reagents and performance of experimental procedures were as described (1)(2)(3)(4)(5)(6). Antibodies were purchased from Cell Signaling Technology (Danvers, MA); Abgent (San Diego, CA); Novus Biologicals (Centennial, CO); Abcam (Cambridge, UK); and Santa Cruz Biotechnology (Dallas, TX).…”

“…Introduction GZ17-6.02 has three components: curcumin, harmine and isovanillin and is presently undergoing phase I safety evaluation in cancer patients with solid tumors and lymphoma (NCT03775525) (1)(2)(3)(4)(5)(6). Over the past two years we have published data demonstrating that GZ17-6.02 kills a wide range of tumor cell types, including ER+ breast, colorectal, pancreatic, hepatic, biliary, NSCLC, cutaneous melanoma, sarcoma and actinic keratoses (1)(2)(3)(4)(5)(6).…”

“…GZ17-6.02 activated PKR-like endoplasmic reticulum kinase (PERK) and increased the phosphorylation (inactivation) at serine 51 of eIF2a, with ER stress signaling facilitating the autophagy response. The mechanisms of cell killing were multi-faceted requiring autophagy, ER stress signaling and death receptor signaling (1)(2)(3)(4)(5)(6).…”

Silencing of lncRNA CRNDE attenuates nonsmall‐cell lung cancer progression by mediating the miR‐455‐3p/HDAC2 axis

“…Therefore, our study was consistent with this previous research. In addition, HDAC2 upregulation could lead to the development of EGFR‐mutant NSCLC 38 . Thus, we could suggest that CRNDE could inhibit the malignant behavior of EGFR‐mutant NSCLC cells by indirectly targeting HDAC2.…”