論文の公開元へIn vivo induction of activin A-producing alveolar macrophages supports the progression of lung cell carcinoma
概要
Title
In vivo induction of activin A-producing
alveolar macrophages supports the progression of
lung cell carcinoma
Author(s)
Taniguchi, Seiji; Matsui, Takahiro; Kimura,
Kenji et al.
Citation
Nature Communications. 2023, 14(1), p. 143
Version Type VoR
URL
rights
https://hdl.handle.net/11094/93152
This article is licensed under a Creative
Commons Attribution 4.0 International License.
Note
Osaka University Knowledge Archive : OUKA
https://ir.library.osaka-u.ac.jp/
Osaka University
Article
https://doi.org/10.1038/s41467-022-35701-8
In vivo induction of activin A-producing
alveolar macrophages supports the
progression of lung cell carcinoma
Received: 30 March 2022
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Accepted: 16 December 2022
Seiji Taniguchi1,2,3, Takahiro Matsui 1,4 , Kenji Kimura3, Soichiro Funaki3,
Yu Miyamoto1,2, Yutaka Uchida 1,2, Takao Sudo1,2, Junichi Kikuta 1,2,5,
Tetsuya Hara6, Daisuke Motooka7,8, Yu-Chen Liu8, Daisuke Okuzaki 7,8,
Eiichi Morii 4, Noriaki Emoto 6, Yasushi Shintani 3 & Masaru Ishii 1,2,5
Alveolar macrophages (AMs) are crucial for maintaining normal lung function.
They are abundant in lung cancer tissues, but their pathophysiological significance remains unknown. Here we show, using an orthotopic murine lung
cancer model and human carcinoma samples, that AMs support cancer cell
proliferation and thus contribute to unfavourable outcome. Inhibin beta A
(INHBA) expression is upregulated in AMs under tumor-bearing conditions,
leading to the secretion of activin A, a homodimer of INHBA. Accordingly,
follistatin, an antagonist of activin A is able to inhibit lung cancer cell proliferation. Single-cell RNA sequence analysis identifies a characteristic subset
of AMs specifically induced in the tumor environment that are abundant in
INHBA, and distinct from INHBA-expressing AMs in normal lungs. Moreover,
postnatal deletion of INHBA/activin A could limit tumor growth in experimental models. Collectively, our findings demonstrate the critical pathological
role of activin A-producing AMs in tumorigenesis, and provides means to
clearly distinguish them from their healthy counterparts.
Cancerous tissues comprise a wide variety of cells in addition to tumor
cells, such as immune cells, fibroblasts1, endothelial cells2, and neural
cells3, which constitute unique microenvironments specific to the cancer
cell type. In particular, different types of immune cells have been
demonstrated to play critical roles in suppressing or promoting tumor
progression in relation to their environment in vivo. Various chemokines
secreted by cancer cells mobilize immune cells, such as cytotoxic CD8+ T
lymphocytes, NK cells, and dendritic cells that attack tumors, into the
cancer microenvironment4. In contrast, immunosuppressive cell types
are also recruited by tumor-secreting cytokines/chemokines, which
serve as ‘internal enemies’ to promote cancer proliferation, invasion, and
metastasis. For example, myeloid-derived suppressor cells (MDSCs)
and tumor-associated macrophages (TAMs) are mobilized to the
cancer microenvironment via systemic circulation to promote tumor
progression5. Among them, TAMs are the most abundant, and the
majority differentiate from bone marrow-derived Ly6c+ inflammatory
monocytes6–8. TAMs are thought to be influenced by cancer cells after
mobilization to transform into phenotypes that benefit the tumor9.
1
Department of Immunology and Cell Biology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan. 2Laboratory of Immunology and
Cell Biology, WPI-Immunology Frontier Research Center, Osaka University, Suita, Osaka 565-0871, Japan. 3Department of General Thoracic Surgery, Osaka
University Graduate School of Medicine, Suita, Osaka 565-0871, Japan. 4Department of Pathology, Osaka University Graduate School of Medicine, Suita,
Osaka 565-0871, Japan. 5Laboratory of Bioimaging and Drug Discovery, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 5670085, Japan. 6Laboratory of Clinical Pharmaceutical Science, Kobe Pharmaceutical University, Higashinada, Kobe 658-8558, Japan. 7Genome Information
Research Center, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan. 8Laboratory of Human Immunology (Single Cell
e-mail: matsuit@molpath.med.osaka-u.ac.jp;
Genomics), WPI-Immunology Frontier Research Center, Osaka University, Suita, Osaka 565-0871, Japan.
mishii@icb.med.osaka-u.ac.jp
Nature Communications | (2023)14:143
1
Article
For instance, TAMs secrete angiogenic factors such as vascular endothelial growth factor to promote tumor angiogenesis and invasion. They
also produce transforming growth factor-β (TGF-β) and epidermal
growth factor to induce epithelial-mesenchymal transition (EMT) during
tumor metastasis. Additionally, TAMs exhibit immunomodulatory
properties: TAMs can produce IL-10, TGF-β, and prostaglandin E2;
mobilize regulatory T cells (Tregs) via C-C motif chemokine 2 (CCL2);
and express programmed death ligand 1/2 (PD-L1/L2) and CD80/86 (B71/2) on their cell surfaces to inhibit immune effector cell activation10. A
comprehensive understanding of tumor-associated immune cells is a
prerequisite for the ultimate control of cancer.
Recent studies have revealed that macrophages arise from two
distinct lineages, along with the discovery of tissue-resident macrophages (TRMs), which have a different origin from those derived from
bone marrow monocytes. In certain tissues, such as the brain, liver, and
lungs, TRMs originating from hematopoietic progenitors in the yolk
sac at the embryonic stage can maintain themselves in situ by selfrenewal and exhibit several microenvironment-specific phenotypes
and functions11–13. In the lungs, alveolar macrophages (AMs) are TRMs
residing in alveolar spaces and constitute one of the two macrophage
populations in the lungs, along with interstitial macrophages (IMs) that
are mainly of bone marrow origin14. AMs have been shown to clean
lung surfactants and protect against infection in the homeostatic
state15. In terms of lung cancers, although the involvement of TAMs has
been referred16, the possible roles of AMs, even though they are by far
abundant major macrophage subsets in cancerous tissues, have seldom been examined. Recently, critical pathological functions of lung
TRMs have been suggested based on single-cell RNA sequencing analyses of human non-small cell lung carcinoma (NSCLC) lesions; however, the detailed mechanism of AM-cancer interaction and its
clinicopathological relevance remain unclear17.
Most of the studies aiming to elucidate cancer-induced host
reactions have been done in systems involving ectopically inoculated
cancer cells into easily accessible areas, e.g. subcutaneous tissues of
flanks. In spite of its broad usability, the method does not enable us to
examine the actual phenomenon with cancer cells in their unique
microenvironments, including possible interactions with residential
immune cells.
In this study, by employing an orthotopic lung cancer model, in
which cancer cells are surgically implanted into the left lung, we
identify residential AMs producing activin A in lung cancer loci as
critical players in cancer progression. The data obtained in this more
natural experimental model, together with the analytical results arising
from studying human samples, suggest an important and targetable
role of AMs in lung tumorigenesis.
Results
Lung AMs support proliferation of lung cancer cells
Based on extensive analyses of human clinical histopathological samples of normal and cancerous lung tissues, we observed that CD163positive AMs accumulated in clusters in cancerous tissues, whereas
they were rather sparse in normal alveolar areas (Fig. 1a, Supplementary Table 1). The population of macrophages was significantly
increased in cancer tissues compared to those in normal tissues
(Fig.1b). These results led us to hypothesize that AMs play a role in the
lung cancer microenvironment. Next, we tested the effect of the AM
cell line (AMJ2-C11, derived from the C57BL/6 mouse strain). The
number of lung carcinoma cells (Lewis lung carcinoma; LLC, derived
from the C57BL/6 mouse strain) significantly increased in culture with
the AM cell supernatant (Fig.1c); this was associated with a reduced
doubling time for the cancer cells (Supplementary Fig. S1a). These
results suggested that AMs could influence the proliferation of lung
cancer cells via the secreting of soluble factors.
To further analyze the functional roles of lung AMs in cancer
proliferation in vivo, we examined an original murine orthotopic lung
Nature Communications | (2023)14:143
https://doi.org/10.1038/s41467-022-35701-8
cancer model. LLC cells stably expressing tdTomato fluorescence were
directly inoculated in the left lung (Supplementary Fig. S1b). Lung AMs
have been reported to be CD45hi, autofluorescence+, CD11c+, CD11b−,
Siglec-F+, and F4/80+ 18. We detected a characteristic CD45+ population
with strong autofluorescence emission at 600/60 nm wavelength,
specifically observed in the lung, but not in the bone marrow or blood
(Supplementary Fig. S1c); we defined this population as lung AMs
because of their F4/80+, Siglec-F+, CD11b−, and CD11c+ characteristics
(Fig. 1d and Supplementary Fig. S1d)19. Furthermore, lung CD45+
autofluorescence+ population in LLC-tdTomato-inoculated conditions
included not only Siglec-F+ F4/80+ AMs but also contained a Siglec-F−,
F4/80+, CD11b+ population, which could be considered to be TAMs in
the lung (Fig. 1d and Supplementary Fig. S1e)20. Next, we intratracheally
administrated clodronate liposome (CDL), the reagent for macrophage specific depletion21, to the mice. CD45+ autofluorescence+
lung AMs were lost in CDL-treated mice under saline-inoculated conditions, but CD45+ autofluorescence+ TAMs were retained in tumorbearing conditions, confirming the specific depletion of lung AMs
in CDL-treated mice (Fig. 1d). ...
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