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Sains Malaysiana 55(8)(2026): 1350-1362
http://doi.org/10.17576/jsm-2026-5508-10
Single-Cell Sequencing Exploration of Extrinsic Adaptive Resistance to
PD-1 Inhibitors: Theory of Acupuncture-Associated Intervention in Advanced Non-Small
Cell Lung Cancer
(Penerokaan Penjujukan Sel Tunggal Rintangan Adaptif Ekstrinsik terhadap
Perencat PD-1: Teori Intervensi Berkaitan Akupunktur dalam Kanser Paru-paru
Bukan Sel Kecil Tahap Teruk)
CHENTENG JIANG & ZHIWEI ZANG*
Yantai Hospital of
Traditional Chinese Medicine, Yantai, 264001, China
Diserahkan: 12 Mei 2026/Diterima: 17 Ogos 2026
Abstract
PD-1 and PD-L1 are
respectively the receptor-ligand molecules on T cells and their target cells.
The interaction between them can mediate immunosuppressive signals and induce
immune escape of tumors. PD-1/PD-L1 inhibitors have revolutionized advanced non-small cell lung cancer (NSCLC) treatment, both primary
resistance and acquired resistance persist due to the immunosuppressive tumor
microenvironment and alternative immune checkpoints. Acupuncture shows
immunomodulatory potential, but its synergy with PD-1 blockade remains unclear.
To investigate the underlying mechanisms, we examined single-cell RNA
sequencing profiles derived from NSCLC patients prior to and following PD-1
inhibitor treatment, utilizing the GSE207422 dataset. After quality control,
53,360 cells were retained. Cell types were annotated via unsupervised
clustering, all cell types and T cell subsets were clustered, and T cells differentiation
trajectories were reconstructed. Intercellular communication was assessed with
CellChat, and a mechanistic framework for acupuncture’s synergistic effects was
integrated from the literature. PD-1 inhibitor treatment remodeled the tumor
microenvironment: T cells increased from 23.9% to 49.0%, while tumor cells
decreased. CD8+ cytotoxic T cells expanded from 32.4% to 55.7% of T cells,
whereas CD4+ naïve T cells declined from 67.1% to 42.7%. Pseudotime analysis
indicated differentiation from CD4+ naïve toward CD8+ cytotoxic and CD8+ memory
T cells. CellChat showed extensive T cell–myeloid interactions. Notably
CD99-CD99 was highly expressed between T cells and tumor cells, suggesting an
adaptive resistance mechanism. PD-1 inhibitors drive potent anti-tumor immunity
in NSCLC, marked by CD8+ T cells expansion and tumor cells reduction, but tumor
cells concurrently upregulate CD99-mediated compensatory suppression. We
propose that acupuncture may synergize by enhancing T cell functional fitness,
enabling T cells to resist CD99-PILRα inhibitory signaling, offering a
promising strategy to overcome immunotherapy resistance.
Keywords:
Acupuncture; immune checkpoint; non-small cell lung cancer; PD-1 inhibitor; single-cell
RNA sequencing
Abstrak
PD-1 dan PD-L1
masing-masing merupakan molekul reseptor-ligan pada sel T dan sel sasarannya.
Interaksi antara mereka boleh menjadi perantara isyarat imunosupresif dan
mendorong pelepasan imun tumor. Perencat PD-1/PD-L1 telah merevolusikan rawatan
kanser paru-paru bukan sel kecil (NSCLC) tahap teruk, kedua-dua rintangan
primer dan rintangan yang diperoleh berterusan disebabkan oleh persekitaran mikro
tumor imunosupresif dan titik semak imun alternatif. Akupunktur menunjukkan
potensi imunomodulatori, tetapi sinerginya dengan sekatan PD-1 masih tidak
jelas. Untuk mengkaji mekanisme yang mendasari, kami memeriksa profil
penjujukan RNA sel tunggal yang diperoleh daripada pesakit NSCLC sebelum dan
selepas rawatan perencat PD-1, menggunakan set data GSE207422. Selepas kawalan
kualiti, 53,360 sel dikekalkan. Jenis sel telah dianotasi melalui pengelompokan
tanpa pengawasan, semua jenis sel dan subset sel T telah dikelompokkan dan
trajektori pembezaan sel T telah dibina semula. Komunikasi antara sel telah
dinilai dengan CellChat dan rangka kerja mekanistik untuk kesan sinergi
akupunktur telah disepadukan daripada kepustakaan. Rawatan perencat PD-1
mengubah suai persekitaran mikro tumor: Sel T meningkat daripada 23.9% kepada
49.0%, manakala sel tumor menurun. Sel T sitotoksik CD8+ berkembang daripada
32.4% kepada 55.7% sel T, manakala sel T naif CD4+ menurun daripada 67.1% kepada
42.7%. Analisis pseudomasa menunjukkan pembezaan daripada CD4+ naif terhadap
sel T memori sitotoksik CD8+ dan CD8+. CellChat mendedahkan interaksi sel
T-myeloid yang meluas. CD99-CD99 diekspresikan dengan tinggi antara sel T dan
sel tumor, menunjukkan mekanisme rintangan adaptif. Perencat PD-1 memacu
imuniti anti-tumor yang kuat dalam NSCLC, ditandai dengan pengembangan sel T
CD8+ dan pengurangan sel tumor, tetapi sel tumor secara serentak meningkatkan
penindasan pampasan yang dimediasi CD99. Kami mencadangkan bahawa akupunktur boleh
bersinergi dengan meningkatkan kecergasan fungsi sel T, membolehkan sel T
menentang isyarat perencat CD99-PILRα, memberikan strategi yang berpotensi
untuk mengatasi rintangan imunoterapi.
Kata kunci:
Akupunktur; kanser paru-paru bukan sel kecil; penjujukan RNA sel tunggal;
perencat PD-1; pusat pemeriksaan imun
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*Pengarang untuk surat-menyurat; email: 860922902@qq.com
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