Effects of inhalable gene transfection as a novel gene therapy for non-small cell lung cancer and malignant pleural mesothelioma

Our plain-English summary of Effects of inhalable gene transfection as a novel gene therapy for non-small cell lung cancer and malignant pleural mesothelioma, by M. Ichikawa, N. Muramatsu, W. Matsunaga, et al (Scientific Reports, 2022).

The study

• Authors: M. Ichikawa, N. Muramatsu, W. Matsunaga, T. Ishikawa, T. Okuda, H. Okamoto, A. Gotoh
• Journal: Scientific Reports
• Published: 2022;12(1):8634
• Cited by: 12 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Gene therapy using vectors has attracted attention in recent years for the treatment of cancers caused by gene mutations. Besides, new treatments are imperative for lung cancer, including non-small cell lung cancer (NSCLC) and malignant pleural mesothelioma (MPM), due to its high mortality. We developed a minimally invasive and orally inhalable tumor suppressor gene drug (SFD-p16 and SFD-p53) with non-viral vectors for lung cancer treatment by combining tumor suppressor genes with an inhalant powder that can deliver active ingredients directly to the lung. We used NSCLC (A549 and H1299) and MPM (H2052) cell lines in an air-liquid interface culture. Transfection of A549 and H2052 cells with SFD-p16 significantly increased p16 mRNA expression levels and decreased cell proliferation in both cell lines. Similar results were obtained with transfection of H1299 with the inhalable gene drug SFD-p53. In an in vivo experiment, a mouse model of lung cancer with orthotopically transplanted luciferase-expressing A549 cells was subjected to intratracheal insufflation of SFD-p16. Consequently, SFD-p16 effectively and directly affected lung cancer. This study suggests that inhalable gene drugs are effective treatments for NSCLC and MPM. We expect inhalable gene drugs to present a novel gene therapy agent for lung cancer that patients can self-administer.

© 2022. The Author(s).

In plain English

This was a laboratory study in cultured human cancer cells and in mice, testing inhalable dry-powder gene preparations made by spray freeze-drying plasmid DNA with 50 kDa hyaluronic acid and L-phenylalanine (SFD-p16 and SFD-p53), compared with a powder containing no plasmid (SFD-placebo), naked plasmid DNA, and untreated controls. The cells used were the non-small cell lung cancer lines A549 and H1299 and the malignant pleural mesothelioma line H2052, grown in air-liquid interface culture. Twenty-four hours after spraying, p16 expression was significantly higher than control in A549 (p less than 0.01) and H2052 (p = 0.012), and p53 expression was higher in H1299 (p less than 0.01); naked p53 plasmid DNA gave no significant expression. Growth inhibition rates were 49.9% for SFD-p16 in A549 and 36.1% in H2052 (placebo 9.3% and 10.1%), and 52.1% for SFD-p53 in H1299 (placebo 29.8%). In mice, subcutaneous tumours sprayed with SFD-p16 were significantly smaller than those sprayed with SFD-placebo (p = 0.021), and intratracheal SFD-p16 (0.5 mg per mouse) inhibited the increase in lung luminescence compared with no treatment, significantly so on day 7. The authors concluded that inhalable gene drugs may be a novel therapy for NSCLC and MPM.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory experimental study: in vitro cell culture experiments plus ex vivo and in vivo mouse experiments
• Who was studied: Human cell lines A549 and H1299 (non-small cell lung cancer) and H2052 (malignant pleural mesothelioma), and male BALB/c nu/nu nude mice (5- and 6-week-old) with subcutaneous or orthotopic A549 tumours; work done in Japan (Hyogo College of Medicine, Meijo University, StateArt Inc.)
• Exposure: Spray freeze-dried inhalable powders containing plasmid DNA coding human p16INK4a (SFD-p16) or human p53 (SFD-p53), 10 µg plasmid in 0.5 mg powder with 50 kDa hyaluronic acid and L-phenylalanine; compared with SFD-placebo (no plasmid), naked pDNA and untreated controls; mouse dose 0.5 mg per mouse by intratracheal insufflation
• Main results:
  • In A549 and H2052 cells, p16 expression after SFD-p16 was significantly higher than control (A549 p less than 0.01; H2052 p = 0.012); in H1299 cells p53 expression after SFD-p53 was significantly higher than control (p less than 0.01), while naked p53 pDNA showed no significant expression
  • Growth inhibition in A549 cells was 49.9% with SFD-p16 versus 9.3% with SFD-placebo
  • In H2052 mesothelioma cells the SFD-p16 growth inhibition rate was 36.1% compared with 10.1% for control and placebo
  • In H1299 cells both SFD-placebo (29.8%) and SFD-p53 (52.1%) suppressed growth, with SFD-p53 significantly more effective than placebo
  • Ex vivo treatment of A549 cells before subcutaneous transplantation gave no significant difference between SFD-p16 and SFD-placebo
  • In mice with established subcutaneous tumours sprayed with powder, average tumour volume was significantly smaller in the SFD-p16 group than the SFD-placebo group (p = 0.021)
  • In the orthotopic lung cancer mouse model, intratracheal SFD-p16 inhibited the increase in lung luminescence compared with no treatment, significant on day 7, while SFD-placebo did not
• The authors' conclusion: The authors concluded that inhalable gene drugs containing tumour suppressor genes suppressed proliferation of NSCLC and MPM cells more than placebo powder and may offer a novel, minimally invasive gene therapy for these cancers
• Limitations the authors state:
  • The authors stated the mechanism of gene transfer by the inhalable gene drugs was not revealed by this experiment
  • The authors stated they could not rule out that hyaluronic acid itself affected cell growth, as placebo powder inhibited growth of H1299 cells
  • The authors stated that in the ex vivo experiment gene transfer may have been insufficient because the powder dissolved in residual medium
  • The authors reported the aerodynamic diameter measurement of SFD-p16 without presenting data

Topics

The National Library of Medicine indexes this study under subjects our site covers: Lung cancer, Mesothelioma.

Medical terms in this study, explained in our Lexicon: Non-Small Cell Lung Cancer, Pleural Mesothelioma, Lung Cancer, Tumor Suppressor Gene, A549 Cells.

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