bims-meluca 2018-10-14 papers

iScience. 2018 Sep 19. pii: S2589-0042(18)30147-0. [Epub ahead of print]8 74-84

A Potent and Selective ULK1 Inhibitor Suppresses Autophagy and Sensitizes Cancer Cells to Nutrient Stress.

Katie R Martin, Stephanie L Celano, Abigail R Solitro, Hakan Gunaydin, Mark Scott, Ronan C O'Hagan, Stuart D Shumway, Peter Fuller, Jeffrey P MacKeigan.

In response to stress, cancer cells generate nutrients and energy through a cellular recycling process called autophagy, which can promote survival and tumor progression. Accordingly, autophagy inhibition has emerged as a potential cancer treatment strategy. Inhibitors targeting ULK1, an essential and early autophagy regulator, have provided proof of concept for targeting this kinase to inhibit autophagy; however, these are limited individually in their potency, selectivity, or cellular activity. In this study, we report two small molecule ULK1 inhibitors, ULK-100 and ULK-101, and establish superior potency and selectivity over a noteworthy published inhibitor. Moreover, we show that ULK-101 suppresses autophagy induction and autophagic flux in response to different stimuli. Finally, we use ULK-101 to demonstrate that ULK1 inhibition sensitizes KRAS mutant lung cancer cells to nutrient stress. ULK-101 represents a powerful molecular tool to study the role of autophagy in cancer cells and to evaluate the therapeutic potential of autophagy inhibition.

Keywords: Cancer; Functional Aspects of Cell Biology; Therapeutics

DOI: https://doi.org/10.1016/j.isci.2018.09.012

Pharmacol Ther. 2018 Oct 03. pii: S0163-7258(18)30177-3. [Epub ahead of print]

Acetylcholine signaling system in progression of lung cancers.

Jamie R Friedman, Stephen D Richbart, Justin C Merritt, Kathleen C Brown, Nicholas A Nolan, Austin T Akers, Jamie K Lau, Zachary R Robateau, Sarah L Miles, Piyali Dasgupta.

The neurotransmitter acetylcholine (ACh) acts as an autocrine growth factor for human lung cancer. Several lines of evidence show that lung cancer cells express all of the proteins required for the uptake of choline (choline transporter 1, choline transporter-like proteins) synthesis of ACh (choline acetyltransferase, carnitine acetyltransferase), transport of ACh (vesicular acetylcholine transport, OCTs, OCTNs) and degradation of ACh (acetylcholinesterase, butyrylcholinesterase). The released ACh binds back to nicotinic (nAChRs) and muscarinic receptors on lung cancer cells to accelerate their proliferation, migration and invasion. Out of all components of the cholinergic pathway, the nAChR-signaling has been studied the most intensely. The reason for this trend is due to genome-wide data studies showing that nicotinic receptor subtypes are involved in lung cancer risk, the relationship between cigarette smoke and lung cancer risk as well as the rising popularity of electronic cigarettes considered by many as a "safe" alternative to smoking. There are a small number of review articles which review the contribution of the other cholinergic proteins in the pathophysiology of lung cancer. The primary objective of this review article is to discuss the function of the acetylcholine-signaling proteins in the progression of lung cancer. The investigation of the role of cholinergic network in lung cancer will pave the way to novel molecular targets and drugs in this lethal malignancy.

Keywords: Acetylcholine; Anti-cancer drugs; Cholinergic; Invasion; Lung cancer; Proliferation

DOI: https://doi.org/10.1016/j.pharmthera.2018.10.002

J Pharm Biomed Anal. 2018 Sep 28. pii: S0731-7085(18)31696-0. [Epub ahead of print]163 122-129

Metabolomic study of serum, urine and bronchoalveolar lavage fluid based on gas chromatography mass spectrometry to delve into the pathology of lung cancer.

Belén Callejón-Leblic, Tamara García-Barrera, Antonio Pereira-Vega, José Luis Gómez-Ariza.

This study explores for the first time the combination of serum, urine and bronchoalveolar lavage fluid (BALF) to deep insight into the pathology of lung cancer (LC) using a metabolomic platform based on gas chromatography mass spectrometry (GCMS). The study includes LC patients, healthy control group (HC) and a group of patients with noncancerous lung diseases (NCC) used as a control group respect to BALF because of the invasive nature this fluid collection. The metabolomic platform was applied to serum, urine and BALF samples in order to compare the metabolomic profiles of these biological fluids and establish metabolic similarities and differences between them. The application of PLS-DA presented a clear classification of groups for all types of samples, indicating the existence of altered metabolites in LC. Twenty six and thirty one perturbed metabolites in the LC were annotated in the comparison of serum and urine samples. On the other hand, sixteen metabolites were altered in BALF of LC patients compared to NCC. The pathway analysis indicated that several amino acid metabolic routes were the most affected in LC. Finally, ROC curves were applied to the dataset and metabolites with an AUC value higher than 0.75 were considered as relevant in the progression of LC.

Keywords: Bronchoalveolar lavage; Gas chromatography; Lung cancer; Metabolomics; Serum; Urine

DOI: https://doi.org/10.1016/j.jpba.2018.09.055

Anal Chim Acta. 2018 Dec 11. pii: S0003-2670(18)30279-4. [Epub ahead of print]1037 256-264

Exosomal lipids for classifying early and late stage non-small cell lung cancer.

Teresa W M Fan, Xiaofei Zhang, Chi Wang, Ye Yang, Woo-Young Kang, Susanne Arnold, Richard M Higashi, Jinze Liu, Andrew N Lane.

Lung cancer is the leading cause of cancer deaths in the United States. Patients with early stage lung cancer have the best prognosis with surgical removal of the tumor, but the disease is often asymptomatic until advanced disease develops, and there are no effective blood-based screening methods for early detection of lung cancer in at-risk populations. We have explored the lipid profiles of blood plasma exosomes using ultra high-resolution Fourier transform mass spectrometry (UHR-FTMS) for early detection of the prevalent non-small cell lung cancers (NSCLC). Exosomes are nanovehicles released by various cells and tumor tissues to elicit important biofunctions such as immune modulation and tumor development. Plasma exosomal lipid profiles were acquired from 39 normal and 91 NSCLC subjects (44 early stage and 47 late stage). We have applied two multivariate statistical methods, Random Forest (RF) and Least Absolute Shrinkage and Selection Operator (LASSO) to classify the data. For the RF method, the Gini importance of the assigned lipids was calculated to select 16 lipids with top importance. Using the LASSO method, 7 features were selected based on a grouped LASSO penalty. The Area Under the Receiver Operating Characteristic curve for early and late stage cancer versus normal subjects using the selected lipid features was 0.85 and 0.88 for RF and 0.79 and 0.77 for LASSO, respectively. These results show the value of RF and LASSO for metabolomics data-based biomarker development, which provide robust an independent classifiers with sparse data sets. Application of LASSO and Random Forests identifies lipid features that successfully distinguish early stage lung cancer patient from healthy individuals.

Keywords: Exosomal lipid profiling; LASSO; Non-small cell lung cancer; Random forest; Ultrahigh resolution Fourier transform mass spectrometry

DOI: https://doi.org/10.1016/j.aca.2018.02.051

Pulm Pharmacol Ther. 2018 Oct 03. pii: S1094-5539(18)30126-3. [Epub ahead of print]

Salvianolic acid B as an anti-emphysema agent I: In vitro stimulation of lung cell proliferation and migration, and protection against lung cell death, and in vivo lung STAT3 activation and VEGF elevation.

Sneha Dhapare, Masahiro Sakagami.

Emphysema causes progressive and life-threatening alveolar structural destruction/loss, yet remains irreversible and incurable to date. Impaired vascular endothelial growth factor (VEGF) signaling has been proposed as a new pathogenic mechanism, and if so, VEGF recovery may enable reversal of emphysema. Thus, we hypothesized that salvianolic acid B (Sal-B), a polyphenol in traditional Chinese herbal danshen, is an alveolar structural recovery agent for emphysema by virtue of VEGF stimulation/elevation via activation of Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3), as stimulating lung cell proliferation and migration, and protecting against lung cell death. Using in vitro human lung microvascular endothelial (HMVEC-L) and alveolar epithelial (A549) cell systems, Sal-B was examined for 1) stimulation of cell proliferation by the MTT and BrdU assays; 2) promotion of cell migration by the scratch wound closure assay; 3) protection against emphysema-like induced cell death by the trypan blue exclusion and flow cytometry assays; and 4) mechanistic involvement of JAK2/STAT3/VEGF in these activities. Sal-B was also spray-dosed to the lungs of healthy rats for two weeks to verify the lung's STAT3 activation and VEGF elevation by western blot, as well as the absence of functional and morphological abnormalities. All the in vitro cell-based activities were concentration-dependent. At 25 μM, Sal-B 1) stimulated cell proliferation by 1.4-2.6-fold; 2) promoted migratory cell wound closure by 1.5-1.7-fold; and 3) protected against cell death induced with H2O2 (oxidative stress) and SU5416 (VEGF receptor blockade) by 49-86%. JAK2 and STAT3 inhibitors and VEGF receptor antagonist each opposed these Sal-B's activities by over 65%, suggesting the mechanistic involvement of JAK2/STAT3 activation and VEGF stimulation/elevation. In rats, Sal-B at 0.2 mg/kg enabled 1.9 and 1.5-fold increased STAT3 phosphorylation and VEGF elevation in the lungs, respectively, while causing no functional and morphological abnormalities. Hence, Sal-B was projected to be a new class of anti-emphysema agent capable of reversing alveolar structural destruction/loss via JAK2/STAT3/VEGF-dependent stimulation of lung cell proliferation and migration, and inhibition of induced lung cell death.

Keywords: Cell death; Emphysema; Migration; Proliferation; STAT3 [signal transducer and activator of transcription 3]; Salvianolic acid B (Sal-B); VEGF [vascular endothelial growth factor]

DOI: https://doi.org/10.1016/j.pupt.2018.10.001