SHP2抑製劑(SHP2i)以單獨和各種組合的臨床試驗正在針對RAS/ERK途徑過度激活的多種腫瘤進行。除了潛在的腫瘤細胞自主作用外，SHP2還對腫瘤微環境(TME)具有重要作用，包括對抗腫瘤免疫的潛在複雜作用。SHP2i的大多數臨床前研究都使用了缺乏適應性免疫反應或很少具有同源人類疾病突變譜且無法複製組織特異性免疫的模型。為此，更好地反映人類癌症的原位和免疫活性腫瘤模型，可能給鑒定增強SHP2i免疫調節作用的更有效、合理的組合策略，提供重要見解。
Clinical trials of SHP2 inhibitors (SHP2i) alone and in various combinations are ongoing for multiple tumors with over-activation of the RAS/ERK pathway. In addition to its potential tumor cell-autonomous actions, SHP2is have important effects on the tumor microenvironment (TME), including potentially complex effects on anti-tumor immunity. Most pre-clinical studies of SHP2is have used models lacking adaptive immune responses or rarely harbor the mutational spectrum of the cognate human disease and fail to reproduce tissue-specific immunity. To this end, orthotopic and immune-competent tumor models that better reflect human cancers might provide important insights into identifying more efficacious and rational combinational strategies that enhance immune-modulatory effects of SHP2is.
方法：為了表征SHP2抑製的腫瘤細胞自主和非自主效應，我們在基因工程小鼠模型(GEMM)中進行了係統的TME分析，包括多色流式細胞術，免疫細胞耗竭實驗和單細胞RNA-Seq。Kras-和Egfr-突變的非小細胞肺癌(NSCLC)連續磁共振成像(MRI)用於評估合理組合的功效，其逆轉SHP2抑製對腫瘤相關免疫細胞的不利後果。此外，我們對來自KRASG12C抑製劑臨床試驗的治療前和治療後活組織檢查樣品進行RNA-seq，以觀察其他RAS/ERK途徑抑製是否在非小細胞肺癌患者中導致相同的不良免疫效應。
Methods:In order to characterize the tumor cell-autonomous and non-autonomous effects of SHP2 inhibition, we conducted systematic TME analysis including multi-color flow cytometry, immune cell depletion experiments and single cell RNA-Seq in genetically engineered mouse models (GEMMs) ofKras-andEgfr-mutant non-small cell lung cancer (NSCLC). Consecutive magnetic resonance imaging (MRI) was used to evaluate the efficacy of a rational combination which reversed the adverse consequences of SHP2 inhibition on tumor-associated immune cells. Moreover, we performed RNA-seq on pre- and post-treatment biopsy samples from KRASG12Cinhibitor clinical trials to see whether other RAS/ERK pathway inhibition results in the same adverse immune effects in NSCLC patients.
結果：我們發現SHP2i治療耗盡肺泡和M2樣巨噬細胞，並促進Kras和Egfr突變NSCLC中的B和T淋巴細胞浸潤。然而，治療還通過CXCR2配體的腫瘤內在的NF-kB依賴性產生增加腫瘤內粒細胞源性抑製細胞(GMDSC)。其他RAS/ERK途徑抑製劑也在小鼠中誘導CXCR2配體和gMDSC流入，並且在KRASG12C抑製劑試驗的患者的腫瘤中誘導CXCR2配體。組合的SHP2(SHP099)/CXCR1/2(SX682)抑製耗盡了S100a8/9高GMDSC的特定簇，產生具有強細胞毒性表型但也表達檢查點受體NKG2A的Klrg1+CD8+效應T細胞，並且在Kras中增強存活率。
Results:We found that SHP2i treatment depleted alveolar and M2-like macrophages and promoted B and T lymphocyte infiltration in Kras- and Egfr-mutant NSCLC. However, treatment also increased intratumoral granulocytic myeloid-derived suppressor cells (gMDSCs) via tumor-intrinsic, NF-kB dependent production of CXCR2 ligands. Other RAS/ERK pathway inhibitors also induced CXCR2 ligands and gMDSC influx in mice, and CXCR2 ligands were induced in tumors from patients on KRASG12C-inhibitor trials. Combined SHP2 (SHP099)/CXCR1/2(SX682) inhibition depleted a specific cluster ofS100a8/9highgMDSCs, generatedKlrg1+ CD8+ effector T cells with a strong cytotoxic phenotype but with also expressing the checkpoint receptor NKG2A, and enhanced survival inKras-andEgfr-mutant models.
結論：研究表明抑製SHP2/RAS/ERK途徑觸發NF-κB依賴性CXCR2配體的上調和S100a8/9高GMDSC的募集，其抑製NSCLC中的T細胞。結合SHP2和CXCR2抑製劑阻斷這種gMDSC移入，導致Th1極化增強，誘導具有高細胞毒活性的CD8+KLRG1+效應T細胞並改善多種NSCLC模型中的存活。實驗結果主張在NSCLC患者中測試RAS/ERK途徑/CXCR1/2/NKG2A抑製劑組合。
Conclusion:Our study shows that inhibiting the SHP2/RAS/ERK pathway triggers NF-kB-dependent upregulation of CXCR2 ligands and recruitment ofS100a8/9highgMDSCs, which suppress T cells in NSCLC. Combining SHP2 and CXCR2 inhibitors blocks this gMDSC immigration, resulting in enhanced Th1 polarization, induction of CD8+ KLRG1+ effector T cells with high cytotoxic activity and improved survival in multiple NSCLC models. Our results argue for testing RAS/ERK pathway/CXCR1/2/NKG2A inhibitor combinations in NSCLC patients.
