研究背景: 腫瘤微環境中的癌細胞與免疫細胞交互作用參與癌症的
進程。我們先前研究結果顯示在初期的癌症上皮細胞中，Rab37 調控
游離的白血球介素1 受體樣1 (soluble interleukin 1 receptor like 1,
ILIRL1, sST2) 的胞吐作用 (exocytosis)，使巨噬細胞較易趨化為M1
型態，進而抑制癌症的生長。值得注意的是白血球介素33 (interleukin
33, IL-33) 會與巨噬細胞膜上ILIRL1 的穿膜蛋白 (long form
transmembrane ST2, ST2L) 結合，活化下游訊號傳遞並促進M2 巨噬
細胞的極化，導致促腫瘤生長的微環境。然而，巨噬細胞中的Rab37
是否調控ST2L 的胞外運輸角色仍然不清楚。
研究目的: 因此，本研究探討於肺癌進程中，Rab37 在巨噬細胞中調
控ST2L 的胞吐作用至細胞膜的機制和角色。
研究結果: 根據共聚焦顯微鏡的免疫螢光染色 (immunofluorescence)
結果顯示，Rab37 與ST2L 在巨噬細胞有共定位 (colocalization) 的現
象，所分析的巨噬細胞包括RAW264.7、THP-1、永生的骨髓源性巨
噬細胞immortalized bone marrow derived macrophages (iBMs) 與老鼠
之骨髓源性巨噬細胞 (bone marrow-derived macrophages，BMDMs)。
為了探討Rab37 是否介導ST2L 的胞內運輸，我們在老鼠巨噬細胞株
RAW264.7 之囊泡分離 (vesicle isolation) 實驗中，證明ST2L 的確存
在於Rab37 所調控的囊泡中；重要的是巨噬細胞RAW264.7 中Rab37
以鳥苷核苷酸依賴性的方式 (GTP-dependent manner) 調控ST2L 的
穿膜表現，此ST2L 會與重組的白血球介素33 (interleukin 33, IL-33)
結合，調控NF-κB、p-38 與JNK 的磷酸化，並促進NF-B 的入核，
上述實驗結果確認巨噬細胞中由Rab37 所運送的ST2L 具有功能性。
為了探討ST2L 是否適合作為治療性靶標，我們開發了中和ST2L 及
IL-33 的抗體藥物，實驗證實ST2L 及IL-33 的中和抗體藥物可抑制老
鼠之骨髓源性巨噬細胞 (BMDMs) 中M2 的極化。藉由肺癌病患癌組
織的免疫組織化學分析 (immunohistochemistry analysis)，我們發現浸
潤的M2 巨噬細胞中Rab37 與ST2L 之間的相關性相較於初期的肺癌
病人，在末期的肺癌病人有更高的相關性。
研究結論：我們的結果顯示Rab37 介導ST2L 的穿膜運輸，Rab37/ST2L
路徑也趨化巨噬細胞走向M2 促腫瘤生長的型態，腫瘤浸潤M2 巨噬
細胞共定位Rab37 及ST2 可能為臨床惡化的證據。

Background: Macrophages in the tumor microenvironment play
important roles in modulating tumor growth and metastasis. In our
previous study, Rab37 small GTPase in early stage cancer epithelial cells,
functions as a modulator of macrophage polarization via regulating the
exocytosis of soluble interleukin 1 receptor like 1 (soluble IL1RL1, also
named as sST2), resulting in the preferential shift of macrophages
phenotype from M2-like to M1-like, and thereby inhibiting tumor growth.
Notably, IL-33 binds to transmembrane form of ILIRL1 (also named
ST2L) that are generally located on the plasma membrane (PM) of
macrophages to activate downstream signaling and skews polarization of
M2 macrophages leading to pro-tumor microenvironment. However, the
role of Rab37 in trafficking of ST2L in macrophages is not well
understood.
Purpose: Our aims are to investigate the role and mechanism of Rab37 in
trafficking of ST2L on the cell membrane of macrophage in promoting
lung cancer progression.
Results: We found colocalization of Rab37 and ST2L in an
IL-33-dependent manner in macrophage cell lines RAW264.7, THP-1,
and immortalized bone marrow derived macrophages (iBMs) and ex vivo
isolated bone marrow derived macrophages (BMDMs) by confocal
microscopy. To investigate whether Rab37 was involved in ST2L
intracellular trafficking, vesicle isolation results showed the presence of
ST2L in Rab37-specific vesicles. Importantly, we found that the level of
ST2L in PM presentation on RAW264.7 cells was more in Rab37
wild-type (WT) and constitutively active form (Q89L) expressing cells
than that in control and dominant negative form (T43N) expression
RAW264.7 cells. Furthermore, to confirm the function of ST2L mediated
by Rab37, recombinant IL-33 binding with ST2L induced NF-κB nuclear
translocation and activation of NF-κB, p-38, and JNK phosphorylation in
a Rab37 GTPase-dependent manner in RAW264.7 cells. To validate
ST2L as a therapeutic target, we developed anti-IL33 and anti-ST2L
neutralized antibodies. We found that neutralizing IL-33 or ST2L
antibody inhibited M2 polarization in BMDMs in vitro. Finally, we
performed immunohistochemistry analysis on tumor specimens from lung
cancer patients to show that the correlation between Rab37 and ST2L in
tumor infiltrated M2 macrophages were more in tumors derived from
advanced stage lung cancer patients than those from early staged patients.
Conclusion: Our results provide first trafficking mode of ST2L mediated
by Rab37 small GTPase and novel evidence of the tumor promoting
function of Rab37/ST2L axis in M2 macrophage and its clinical
implications.

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