Labyrinthin Expression Is Associated with Poor Prognosis in Patients with Non-Small-Cell Lung Cancer

Weijie Ma, Jie Zeng, Dennis J Montoya, Kyra Toomey, Chihong Zhou, Shuai Chen, Dingning Liu, Michael Babich, James A Radosevich, Tianhong Li, Weijie Ma, Jie Zeng, Dennis J Montoya, Kyra Toomey, Chihong Zhou, Shuai Chen, Dingning Liu, Michael Babich, James A Radosevich, Tianhong Li

Abstract

To determine Labyrinthin (LAB) expression in non-small-cell lung cancer (NSCLC), we immunostained and scored for LAB immunohistochemistry (IHC) expression on sections of tissue microarrays (TMAs) prepared from 256 archival tissue blocks of NSCLC. Propensity-score-weighted Kaplan-Meier curves and weighted Cox models were used to associate LAB expression with overall survival. LAB mRNA expression was assessed in The Cancer Genome Atlas (TCGA) and correlated with clinical phenotype and outcome. Positive LAB IHC expression (>5% of tumor cells) was detected in 208/256 (81.3%) of NSCLC samples, and found in both lung adenocarcinomas (LUAD) and lung squamous cell cancer (LUSC). LAB positivity was associated with poor overall survival (HR = 3.56, 95% CI: 2.3-5.4; p < 0.0001) and high tumor differentiation grade or metastasis compared with negative LAB expression. Univariant and multivariate survival analyses demonstrated LAB expression as an independent prognostic factor for NSCLC patients. LAB RNA expression in TCGA-LUAD was higher in primary and advanced-stage tumors than in normal tissue, and was associated with poorer overall survival. No significant differences or associations were found with LAB RNA expression in TCGA-LUSC. The LAB IHC assay is being used to identify candidate cancer patients for the first-in-human phase I trial evaluating the LAB vaccines (UCDCC#296, NCT051013560).

Trial registration: ClinicalTrials.gov NCT51013560.

Keywords: RNA sequencing; TCGA; labyrinthin; lung adenocarcinoma; lung squamous cell carcinoma; non-small-cell lung cancer; prognosis; tissue microarray.

Conflict of interest statement

Michael Babich and James A. Radosevich are principals and shareholders in LabyRx Immuno-Oncology, Inc. They have patents related to labyrinthin. Tianhong Li has received research grants from LabyRx Immuno-Oncology, Inc. Other authors declare that they have no competing interest.

Figures

Figure 1
Figure 1
Study flow chart. The distribution of lung cancer types of TMA is illustrated. Abbreviations: NSCLC, non-small-cell lung cancer; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; NOS, not other specified.
Figure 2
Figure 2
Atlas of LAB expression by IHC. (A) Representative pictures of LAB stain in normal lung tissue. (B–E) Atlas of representative pictures of 0, 1, 2, and 3 IHC scores for LAB expression in NSCLC TMAs. Abbreviations: LAB, labyrinthin; IHC, immunohistochemistry; NSCLC, non-small-cell lung cancer; TMA, tissue microarray; TPS, tumor proportion score.
Figure 3
Figure 3
LAB expression and NSCLC metastasis. (A) Increased LAB expression was detected in NSCLC compared to matched, non-cancerous tissues. (B) Increased LAB expression was detected in LUAD and LUSC compared to matched, non-cancerous tissues, and brain metastasis. (C) Higher LAB expression was associated with NSCLC metastasis. ** p < 0.01 and *** p < 0.001 for statistical significance.
Figure 4
Figure 4
High LAB expression was associated with poor prognosis in NSCLC patients. (A) Kaplan–Meier curves of overall survival were stratified by LAB expression score (0, 1, 2, and 3). (B) Kaplan–Meier curves of overall survival were stratified by LAB expression (negative vs. positive).
Figure 5
Figure 5
LAB expression in histology subtypes of NSCLC. (A) The median LAB expression was higher in LUSC than LUAD. Kaplan–Meier curves of overall survival were stratified by LAB expression (negative vs. positive) in LUAD (B) and LUSC (C), respectively.
Figure 6
Figure 6
LAB RNA expression in TCGA. Average LAB exon RNA expression was assessed in the TCGA LUAD. Violin plots of average LAB expression in LUAD by tissue type (A) or cancer stage (B). Pairwise statistical differences assessed by Wilcoxon rank sum test (Mann–Whitney U test) and one-way ANOVA by Kruskal–Wallis tests. (C) Kaplan–Meier curves depicting overall survival or patients with higher median LAB RNA expression. Number at risk table shown below. Log-rank test used to evaluate significance. Same analyses applied to TCGA LUSC (D–F). * p < 0.05 and *** p < 0.001 for statistical significance. ns for no statistical significance.

References

    1. Babich M., Sharma A., Li T., Radosevich J.A. Labyrinthin: A distinct pan-adenocarcinoma diagnostic and immunotherapeutic tumor specific antigen. Heliyon. 2022;8:e08988. doi: 10.1016/j.heliyon.2022.e08988.
    1. Zheng W., Wang X., Hu J., Bai B., Zhu H. Diverse molecular functions of aspartate β-hydroxylase in cancer (Review) Oncol. Rep. 2020;44:2364–2372. doi: 10.3892/or.2020.7792.
    1. Combs S., Radosevich J., Ma Y., Lee I., Gould V., Battifora H., Rosen S. Expression of the Antigenic Determinant Recognized by the Monoclonal Antibody 44-3A6 on Select Human Adenocarcinomas and Normal Human Tissues. Tumor Biol. 1988;9:116–122. doi: 10.1159/000217552.
    1. Siddiqui F., Iqbal Z., Radosevich J. Changes in the Expression of the Tumor-Associated Antigen Recognized by Monoclonal Antibody 44–3A6 in A549 Cells due to Calcium. Tumor Biol. 1992;13:142–151. doi: 10.1159/000217758.
    1. Padar S., Van Breemen C., Thomas D.W., Uchizono J., Livesey J.C., Rahimian R. Differential regulation of calcium homeostasis in adenocarcinoma cell line A549 and its Taxol-resistant subclone. Br. J. Pharmacol. 2004;142:305–316. doi: 10.1038/sj.bjp.0705755.
    1. Cajulis R.S., Szumel R., Frias-Hidvegi D., Combs S.G., Radosevich J.A. Monoclonal antibody 44-3A6 as an adjunct in cytodiagnosis of adenocarcinomas in body fluids. Diagn. Cytopathol. 1993;9:179–183. doi: 10.1002/dc.2840090214.
    1. Spagnolo D.V., Whitaker D., Carrello S., Radosevich J.A., Rosen S.T., Gould V.E. The Use of Monoclonal Antibody 44-3A6 in Cell Blocks in the Diagnosis of Lung Carcinoma, Carcinomas Metastatic to Lung and Pleura, and Pleural Malignant Mesothelioma. Am. J. Clin. Pathol. 1991;95:322–329. doi: 10.1093/ajcp/95.3.322.
    1. Ma W., Chen S., Ogihara N.L., Huynh J., Yen A., Kelly K., Lara P.N., Radosevich J.A., Babich M., Li T. A first-in-human, phase I, open-label study of a novel cancer vaccine labvax 3(22)-23 and adjuvant GM-CSF in patients with advanced stage adenocarcinomas. J. Clin. Oncol. 2022;40:TPS2700. doi: 10.1200/JCO.2022.40.16_suppl.TPS2700.
    1. Reck M., Rabe K.F. Precision Diagnosis and Treatment for Advanced Non–Small-Cell Lung Cancer. N. Engl. J. Med. 2017;377:849–861. doi: 10.1056/NEJMra1703413.
    1. Postmus P.E., Kerr K.M., Oudkerk M., Senan S., Waller D.A., Vansteenkiste J., Escriu C., Peters S., ESMO Guidelines Committee Early and Locally Advanced Non-Small-Cell Lung Cancer (NSCLC): ESMO Clinical Practice Guidelines for Diagnosis, Treatment and Follow-up. Ann. Oncol. 2017;28:iv1–iv21. doi: 10.1093/annonc/mdx222.
    1. Knight S.B., Phil A., Crosbie P.A., Balata H., Chudziak J., Hussell T., Dive C. Progress and prospects of early detection in lung cancer. Open Biol. 2017;7:170070. doi: 10.1098/rsob.170070.
    1. Tan W.-L., Jain A., Takano A., Newell E.W., Iyer N.G., Lim W.-T., Tan E.-H., Zhai W., Hillmer A.M., Tam W.L., et al. Novel therapeutic targets on the horizon for lung cancer. Lancet Oncol. 2016;17:e347–e362. doi: 10.1016/S1470-2045(16)30123-1.
    1. Ullah A., Leong S.W., Wang J., Wu Q., Ghauri M.A., Sarwar A., Su Q., Zhang Y. Cephalomannine inhibits hypoxia-induced cellular function via the suppression of APEX1/HIF-1α interaction in lung cancer. Cell Death Dis. 2021;12:490. doi: 10.1038/s41419-021-03771-z.
    1. Iqbal H., Menaa F., Khan N.U., Razzaq A., Khan Z.U., Ullah K., Kamal R., Sohail M., Thiripuranathar G., Uzair B., et al. Two Promising Anti-Cancer Compounds, 2-Hydroxycinnaldehyde and 2- Benzoyloxycinnamaldehyde: Where do we stand? Comb. Chem. High Throughput Screen. 2022;25:808–818. doi: 10.2174/1386207324666210216094428.
    1. Ullah A., Aziz T., Ullah N., Nawaz T. Molecular mechanisms of Sanguinarine in cancer prevention and treatment. Anti Cancer Agents Med. Chem. 2022;22:3. doi: 10.2174/1871520622666220831124321.
    1. Li Q., Ma W., Chen S., Tian E.C., Wei S., Fan R.R., Wang T., Zhou C., Li T. High integrin α3 expression is associated with poor prognosis in patients with non-small cell lung cancer. Transl. Lung Cancer Res. 2020;9:1361–1378. doi: 10.21037/tlcr-19-633.
    1. Austin P.C. The performance of different propensity score methods for estimating marginal hazard ratios. Stat. Med. 2013;32:2837–2849. doi: 10.1002/sim.5705.
    1. Imbens G. The role of the propensity score in estimating dose-response functions. Biometrika. 2000;87:706–710. doi: 10.1093/biomet/87.3.706.
    1. Rubin D.B. Multiple Imputation for Nonresponse in Surveys. John Wiley & Sons; Hoboken, NJ, USA: 1987.
    1. Sun W., Duan T., Ye P., Chen K., Zhang G., Lai M., Zhang H. TSVdb: A web-tool for TCGA splicing variants analysis. BMC Genom. 2018;19:405. doi: 10.1186/s12864-018-4775-x.
    1. Mortazavi A., Williams B.A., McCue K., Schaeffer L., Wold B. Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat. Methods. 2008;5:621–628. doi: 10.1038/nmeth.1226.
    1. Kassambara A., Kosinski M., Biecek P., Fabian S. Survminer: Drawing Survival Curves Using ‘Ggplot2’ R Package Version 0.4.4. 2019. [(accessed on 11 October 2022)]. Available online: .
    1. Li L., Goedegebuure S., Gillanders W. Preclinical and clinical development of neoantigen vaccines. Ann. Oncol. 2017;28:xii11–xii17. doi: 10.1093/annonc/mdx681.
    1. Ott P.A., Hu Z., Keskin D.B., Shukla S.A., Sun J., Bozym D.J., Zhang W., Luoma A., Giobbie-Hurder A., Peter L., et al. An immunogenic personal neoantigen vaccine for patients with melanoma. Nature. 2017;547:217–221. doi: 10.1038/nature22991. Erratum in Nature 2018, 555, 402.
    1. Schumacher T.N., Schreiber R.D. Neoantigens in cancer immunotherapy. Science. 2015;348:69–74. doi: 10.1126/science.aaa4971.
    1. Blass E., Ott P.A. Advances in the development of personalized neoantigen-based therapeutic cancer vaccines. Nat. Rev. Clin. Oncol. 2021;18:215–229. doi: 10.1038/s41571-020-00460-2.
    1. Luu M., Sabo E., de la Monte S.M., Greaves W., Wang J., Tavares R., Simao L., Wands J.R., Resnick M.B., Wang L. Prognostic value of aspartyl (asparaginyl)-β-hydroxylase/humbug expression in non–small cell lung carcinoma. Hum. Pathol. 2009;40:639–644. doi: 10.1016/j.humpath.2008.11.001.

Source: PubMed

Sponsorer och medarbetare

Medicinska tillstånd

Läkemedelsinterventioner

3
Prenumerera