Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Dec;30(52):e1805007.
doi: 10.1002/adma.201805007. Epub 2018 Nov 2.

Trapping of Lipopolysaccharide to Promote Immunotherapy against Colorectal Cancer and Attenuate Liver Metastasis

Wantong Song  1   2 , Karthik Tiruthani  3 , Ying Wang  1   3 , Limei Shen  1 , Mengying Hu  1 , Oleksandra Dorosheva  3 , Kunyu Qiu  1 , Karina A Kinghorn  1 , Rihe Liu  3   4 , Leaf Huang  1
Affiliations

Trapping of Lipopolysaccharide to Promote Immunotherapy against Colorectal Cancer and Attenuate Liver Metastasis

Wantong Song et al. Adv Mater. 2018 Dec.

Abstract

The development and progression of colorectal cancer (CRC) is closely related to gut microbiome. Here, the impact of lipopolysaccharide (LPS), one of the most prevalent products in the gut microbiome, on CRC immunotherapy is investigated. It is found that LPS is abundant in orthotopic CRC tissue and is associated with low responses to anti-PD-L1 mAb therapy, and clearance of Gram-negative bacteria from the gut using polymyxin B (PmB) or blockade of Toll-like receptor 4 using TAK-242 will both relieve the immunosuppressive microenvironment and boost T-cell infiltration into the CRC tumor. Further, an engineered LPS-targeting fusion protein is designed and its coding sequence is loaded into a lipid-protamine-DNA (LPD) nanoparticle system for selective expression of LPS trap protein and blocking LPS inside the tumor, and this nanotrapping system significantly relieves the immunosuppressive microenvironment and boosts anti-PD-L1 mAb therapy against CRC tumors. This LPS trap system even attenuates CRC liver metastasis when applied, suggesting the importance of blocking LPS in the gut-liver axis. The strategy applied here may provide a useful new way for treating CRC as well as other epithelial cancers that interact with mucosa microbiome.

Keywords: cancer therapy; drug delivery; immunotherapy; metastasis; nanotechnology.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
a) Growth curves of s.c. and orthotopic CT26-FL3 tumors treated with PBS or 100 μg/mouse anti-PD-L1 antibody (n = 5 mice per group). b) CD3-immunofluorescence images of s.c. and orthotopic CT26-FL3 tumors. Yellow dashed line indicates the boundary between intestinal mucosa and the orthotopic tumor. * p < 0.05, comparison of CD3+ ratios between s.c. tumor and orthotopic tumor (n = 12 to 15 region of interest (ROI) from four tumors per group). c) Endotoxin test of s.c. CT26-FL3 tumors and orthotopic CT26-FL3 tumors (n = 5). d) Treatment schedule of PmB on orthotopic CT26-FL3 tumor bearing mice. e) Endotoxin test of orthotopic CT26-FL3 tumors in mice treated with PBS and PmB (n = 5). f) CD3-immunofluorescence images of orthotopic CT26-FL3 tumor in mice treated with PBS or PmB (day 22 after tumor inoculation). *** p < 0.001, comparison of CD3+ ratios between PBS and PmB groups (n = 12 to 15 ROIs from four tumors per group). g) Flow cytometry analysis of orthotopic CT26-FL3 tumors in PBS and PmB treated groups (day 22, n = 4). ns, no significant difference, * p<0.05, ***p<0.001. h) Relative mRNA expression of various cytokines and chemokines in the PBS and PmB treated tumors (on day 22, n = 6). Significant differences were assessed in a using two-way ANOVA with multiple comparisons and in others using t test. Results are presented as mean (SD).
Figure 2.
Figure 2.
a) The schematic of the trimeric LPS trap and multivalent interaction of the LPS trap with the lipid A region of LPS. The red region is the LPS-binding moiety. b) Preparation of LPS trap plasmid (pTrap) loaded LPD nanoparticles. c) LPS trap protein expression in serum, major organs and CT26-FL3 tumor. LPS trap plasmid loaded LPD was given on day 0, and the LPS trap expression was measured using ELISA by targeting the His (6×)-tag engineered at the C-terminus of the LPS trap on day 2 (n = 3). *** p < 0.001. d) Flow cytometry analysis of orthotopic CT26-FL3 tumor after PBS and LPS trap treatment (on day 22, n = 4). ns, no significant difference, ** p<0.01, ***p<0.001. e) CD3-immunofluorescence images of orthotopic CT26-FL3 tumor after LPS trap treatment (day 22). Yellow dashed line indicates the boundary between intestinal mucosa and the orthotopic tumor. *** p < 0.001, comparison of CD3+ ratios between PBS and LPS trap groups (n = 12 to 15 ROIs from four tumors per group). f-g) Relative mRNA expression of various cytokines or chemokines in the PBS and LPS trap treated tumors (n = 6). Significant differences were assessed using t test. Results are presented as mean (SD).
Figure 3.
Figure 3.
a) Treatment scheme of orthotopic CT26-FL3 tumor. b) Growth curves of orthotopic CT26-FL3 tumors in PBS, α-PD-L1, LPS trap plasmid and LPS trap+α-PD-L1 treated groups (n = 6–10 mice per group). c) Representative bioluminescence imaging of tumor burden, applying a maximum luminescence threshold of 2 × 109. d) Kaplan-Meier survival curves; the difference between different groups is significant by log-rank test. e) Flow cytometry analysis of orthotopic CT26-FL3 tumor after various treatments (on day 27, n = 4). f) CD4+ and CD8+ T cell depletion test of the LPS trap+α-PD-L1 treatment (n = 5 mice per group). Rat IgG, α-CD8 or α-CD4 (200 μg/mouse, i.p. injection) together with LPS trap+α-PD-L1 were given on day 15, 18 and 21. ns, no significant difference, *** p<0.001. g) Pathological analyses of liver after various treatments. The regions pointed by blue arrows are metastasis sites. The photos were taken at 10× magnification. h) Treatment scheme and tumor growth curves of CT26-FL3 liver metastatic tumors (n = 5 mice per group). Significant differences were assessed in b, f, h using two-way ANOVA with multiple comparisons and in e using t test. Results are presented as mean (SD).
See this image and copyright information in PMC

Comment in

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Mellman I, Coukos G, Dranoff G, Nature 2011, 480, 480; - PMC - PubMed
    2. Hoos A, Nature reviews. Drug discovery 2016, 15, 235. - PubMed
    1. Kuipers EJ, Grady WM, Lieberman D, Seufferlein T, Sung JJ, Boelens PG, van de Velde CJH, Watanabe T, Nature Reviews Disease Primers 2015, 1, 15065. - PMC - PubMed
    1. Le DT, Durham JN, Smith KN, Wang H, Bartlett BR, Aulakh LK, Lu S, Kemberling H, Wilt C, Luber BS, Wong F, Azad NS, Rucki AA, Laheru D, Donehower R, Zaheer A, Fisher GA, Crocenzi TS, Lee JJ, Greten TF, Duffy AG, Ciombor KK, Eyring AD, Lam BH, Joe A, Kang SP, Holdhoff M, Danilova L, Cope L, Meyer C, Zhou S, Goldberg RM, Armstrong DK, Bever KM, Fader AN, Taube J, Housseau F, Spetzler D, Xiao N, Pardoll DM, Papadopoulos N, Kinzler KW, Eshleman JR, Vogelstein B, Anders RA, Diaz LA, Science (New York, N.Y.) 2017, 357, 409; - PMC - PubMed
    2. Zou W, Wolchok JD, Chen L, Sci Transl Med 2016, 8, 328rv4. - PMC - PubMed
    1. Le DT, Uram JN, Wang H, Bartlett BR, Kemberling H, Eyring AD, Skora AD, Luber BS, Azad NS, Laheru D, Biedrzycki B, Donehower RC, Zaheer A, Fisher GA, Crocenzi TS, Lee JJ, Duffy SM, Goldberg RM, de la Chapelle A, Koshiji M, Bhaijee F, Huebner T, Hruban RH, Wood LD, Cuka N, Pardoll DM, Papadopoulos N, Kinzler KW, Zhou S, Cornish TC, Taube JM, Anders RA, Eshleman JR, Vogelstein B, Diaz LAJ, New England Journal of Medicine 2015, 372, 2509. - PMC - PubMed
    1. Zelenay S, van der Veen Annemarthe G., Böttcher Jan P., Snelgrove Kathryn J., Rogers N, Acton Sophie E., Chakravarty P, Girotti Maria R., Marais R, Quezada Sergio A., Sahai E, Reis e Sousa C, Cell 2015, 162, 1257. - PMC - PubMed

MeSH terms

Substances