A Mechanophenotyping chip for high-throughput detection of metastatic bacteria-infected circulating tumor cells

1. Sepich-Poore, G. D. et al. The microbiome and human cancer. Science 371, 1331 (2021).

   Google Scholar 

2. Riquelme, E. et al. Tumor microbiome diversity and composition influence pancreatic cancer outcomes. Cell 178, 795–806 (2019).

   Google Scholar 

3. Xi, J. et al. Reverse intratumor bacteria-induced gemcitabine resistance with carbon nanozymes for enhanced tumor catalytic-chemo therapy. Nano Today 43, 101395 (2022).

   Google Scholar 

4. Colbert, L. E. et al. Tumor-resident Lactobacillus iners confer chemoradiation resistance through lactate-induced metabolic rewiring. Cancer Cell 41, 1945–1962 (2023).

   Google Scholar 

5. Nejman, D. et al. The human tumor microbiome is composed of tumor type-specific intracellular bacteria. Science 368, 973–980 (2020).

   Google Scholar 

6. Galeano Nino, J. L. et al. Effect of the intratumoral microbiota on spatial and cellular heterogeneity in cancer. Nature 611, 810–817 (2022).

   Google Scholar 

7. Fu, A. et al. Tumor-resident intracellular microbiota promotes metastatic colonization in breast cancer. Cell 185, 1356–1372 (2022).

   Google Scholar 

8. Fu, A., Yao, B., Dong, T. & Cai, S. Emerging roles of intratumor microbiota in cancer metastasis. Trends Cell Biol. 33, 583–593 (2023).

   Google Scholar 

9. Suhail, Y. et al. Systems biology of cancer metastasis. Cell Syst. 9, 109–127 (2019).

   Google Scholar 

10. Lawrence, R., Watters, M., Davies, C. R., Pantel, K. & Lu, Y. J. Circulating tumour cells for early detection of clinically relevant cancer. Nat. Rev. Clin. Oncol. 20, 487–500 (2023).

    Google Scholar 

11. Poudineh, M., Sargent, E. H., Pantel, K. & Kelley, S. O. Profiling circulating tumour cells and other biomarkers of invasive cancers. Nat. Biomed. Eng. 2, 72–84 (2018).

    Google Scholar 

12. Lin, D. et al. Circulating tumor cells: biology and clinical significance. Signal Transduct. Target Ther. 6, 404 (2021).

    Google Scholar 

13. Chen, Y. et al. A supramolecular co-delivery strategy for combined breast cancer treatment and metastasis prevention. Chin. Chem., Lett. 31, 1153–1158 (2020).

    Google Scholar 

14. Mitchell, M. J. & King, M. R. Fluid shear stress sensitizes cancer cells to receptor-mediated apoptosis via trimeric death receptors. N. J. Phys. 15, 015008 (2013).

    Google Scholar 

15. Han, X. et al. Microfluidic cell deformability assay for rapid and efficient kinase screening with the CRISPR-Cas9 system. Angew. Chem. Int. Ed. 55, 8561–8565 (2016).

    Google Scholar 

16. Han, Y. et al. Aptazyme-induced cascade amplification integrated with a volumetric bar-chart chip for highly sensitive detection of aflatoxin B1 and adenosine triphosphate. Analyst 147, 2500–2507 (2022).

    Google Scholar 

17. Wang, Y., Gao, Y. & Song, Y. Microfluidics-based urine biopsy for cancer diagnosis: recent advances and future trends. Chem. Med. Chem. 17, e20220422 (2022).

    Google Scholar 

18. Kwong, G. A. et al. Synthetic biomarkers: a twenty-first century path to early cancer detection. Nat. Rev. Cancer 21, 655–668 (2021).

    Google Scholar 

19. Guo, Q. et al. Deformability based sorting of red blood cells improves diagnostic sensitivity for malaria caused by plasmodium falciparum. Lab Chip 16, 645–654 (2016).

    Google Scholar 

20. Wang, Y. et al. Emerging trends in organ-on-a-chip systems for drug screening. Acta Pharm. Sin. B 13, 2483–2509 (2023).

    Google Scholar 

21. Gao, Y. et al. An enzyme-loaded metal–organic framework-assisted microfluidic platform enables single-cell metabolite analysis. Angew. Chem. Int. Ed. 62, e202302000 (2023).

    Google Scholar 

22. Wu, J. et al. Identifying the phenotypes of tumor-derived extracellular vesicles using size-coded affinity microbeads. J. Am. Chem. Soc. 144, 23483–23491 (2022).

    Google Scholar 

23. Zhang, P. et al. Ultrasensitive detection of circulating exosomes with a 3D-nanopatterned microfluidic chip. Nat. Biomed. Eng. 3, 438–451 (2019).

    Google Scholar 

24. Liu, J. et al. Magnetic-opticaldual functional Janus particles for the detection of metal ions assisted by machine learning. Smart Mol. 1, e20230006 (2023).

    Google Scholar 

25. Tse, H. T. K. et al. Quantitative diagnosis of malignant pleural effusions by single-cell mechanophenotyping. Sci. Transl. Med. 5, 212ra163 (2013).

    Google Scholar 

26. Urbanska, M. et al. A comparison of microfluidic methods for high-throughput cell deformability measurements. Nat. Methods 17, 587–593 (2020).

    Google Scholar 

27. Nematbakhsh, Y. & Lim, C. T. Cell biomechanics and its applications in human disease diagnosis. Acta Mech. Sin. 31, 268–273 (2015).

    Google Scholar 

28. Di Carlo, D. A mechanical biomarker of cell state in medicine. SLAS Technol. 17, 32–42 (2012).

    Google Scholar 

29. Sarioglu, A. F. et al. A microfluidic device for label-free, physical capture of circulating tumor cell clusters. Nat. Methods 12, 685–691 (2015).

    Google Scholar 

30. Au, S. H. et al. Clusters of circulating tumor cells traverse capillary-sized vessels. Proc. Nat. Acad. Sci. USA 113, 4947–4952 (2016).

    Google Scholar 

31. Jia, Y. et al. Microfluidic tandem mechanical sorting system for enhanced cancer stem cell isolation and ingredient screening. Adv. Healthc. Mater. 10, e2100985 (2021).

    Google Scholar 

32. Han, X. et al. Microfluidic cell trap arrays for single hematopoietic stem/progenitor cell behavior analysis. Proteomics 20, e1900223 (2020).

    Google Scholar 

33. Liu, Z. et al. Integrated microfluidic chip for efficient isolation and deformability analysis of circulating tumor cells. Adv. Biosyst. 2, 1800200 (2018).

    Google Scholar 

34. Park, E. S. et al. Continuous flow deformability-based separation of circulating tumor cells using microfluidic ratchets. Small 12, 1909–1919 (2016).

    Google Scholar 

35. Nobes, C. D. & Hall, A. Rho, Rac, and Cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell 81, 53–62 (1995).

    Google Scholar 

36. van Oosten, M. et al. Real-time in vivo imaging of invasive- and biomaterial-associated bacterial infections using fluorescently labelled vancomycin. Nat. Commun. 4, 2584 (2013).

    Google Scholar 

37. Zhang, B. et al. Synthesis of vancomycin fluorescent probes that retain antimicrobial activity, identify Gram-positive bacteria, and detect Gram-negative outer membrane damage. Commun. Biol. 6, 409 (2023).

    Google Scholar 

38. Iida, N. et al. Commensal bacteria control cancer response to therapy by modulating the tumor microenvironment. Science 342, 967–970 (2013).

    Google Scholar 

39. Pushalkar, S. et al. The pancreatic cancer microbiome promotes oncogenesis by induction of innate and adaptive immune suppression. Cancer Discov. 8, 403–416 (2018).

    Google Scholar 

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Author notes

1. These authors contributed equally: Wen Luo, Yanfeng Gao.

2. These authors jointly supervised this work:Tony Y. Hu, Yujun Song.

Authors and Affiliations

1. Department of Gastric and Hernia Surgery, Nanjing Drum Tower Hospital, College of Engineering and Applied Sciences, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing, China

   Wen Luo & Yujun Song

2. School of Medical Imaging, Wannan Medical College, Wuhu, China

   Yanfeng Gao

3. State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, China

   Shujun Feng

4. Department of Laboratory Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing, China

   Bangshun He & Jingjing Li

5. Department of Biochemistry and Molecular Biology, School of Medicine & Holistic Integrative Medicine, Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing, China

   Jingjing Yang

6. Division of Colorectal Surgery, Department of General Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Nanjing University Medical School, Nanjing, China

   Yi Yin

7. Department of Gastric and Hernia Surgery, Nanjing University Medical School Affiliated Drum Tower Hospital, Nanjing, China

   Meng Wang

8. School of Physics, Nanjing University, Nanjing, China

   Bin Xue & Yi Cao

9. School of Biomedical Engineering, Tsinghua Medicine, Tsinghua University, Beijing, China

   Tony Y. Hu

Contributions

The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. Y. Song, J. Yang, and T. Hu supervised the project. B.H., J.L., M.W. and Y.Y. provided the clinical blood samples. Y.S. and W.L. conceptualized the research, designed experiments, analyzed the data, and wrote the manuscript with feedback from all the authors. Y. Gao assisted in the preparation of microfluidic devices. S. Feng assisted in the in vivo assays. B. Xue and Y. Cao assisted in the AFM measurements.

Corresponding authors

Correspondence to Tony Y. Hu or Yujun Song.

Competing interests

Y.S. and W.L. are inventors on a patent application (number of patent application, 202511249032.X) filed by Nanjing University related to this work. The other authors declare no competing interests.

Peer review information

Nature Communications thanks Esmail Pishbin, Chang Liu, and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. A peer review file is available.

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