Cited by 133Nanoparticle-Based Drug DeliveryAdvanced Drug Delivery SystemsGut microbiota and health

The DOI matches its Crossref record; 56 of the 56 references it cites were found in a registry or an index; 11 of the 53 sources its claims cite were checked for support.

Combining gut microbiota modulation and chemotherapy by capecitabine-loaded prebiotic nanoparticle improves colorectal cancer therapy

Open access
Lang, Tianqun, Zhu, Runqi, Zhu, Xiao, Yan, Wenlu, Li, Yu, Zhai, Yihui, Wu, Ting, Huang, Xin, Yin, Qi, Li, Yaping·2023·Nature Communications

Abstract

Colorectal cancer (CRC) therapy efficiency can be influenced by the microbiota in the gastrointestinal tract. Compared with traditional intervention, prebiotics delivery into the gut is a more controllable method for gut microbiota modulatory therapy. Capecitabine (Cap), the first-line chemotherapeutic agent for CRC, lacks a carrier that can prolong its half-life. Here, we construct a Cap-loaded nanoparticle using the prebiotic xylan-stearic acid conjugate (SCXN). The oral administration of SCXN delays the drug clearance in the blood and increases the intra-tumoral Cap concentration in the CRC mouse model. SCXN also facilitates the probiotic proliferation and short chain fatty acid production. Compared with free Cap, SCXN enhances the anti-tumor immunity and increases the tumor inhibition rate from 5.29 to 71.78%. SCXN exhibits good biocompatibility and prolongs the median survival time of CRC mice from 14 to 33.5 d. This prebiotics-based nanoparticle provides a promising CRC treatment by combining gut microbiota modulation and chemotherapy.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

The paper constructs SCXN, an oral capecitabine-loaded nanoparticle made from a xylan–stearic acid conjugate, to combine chemotherapy with gut microbiota modulation for colorectal cancer. In mouse CRC models, SCXN improved drug pharmacokinetics, tumor inhibition, anti-tumor immunity, probiotic abundance, short-chain fatty acid production, and survival compared with free capecitabine.

Key findings

  • SCXN had a capecitabine drug loading of 9.49% and encapsulation efficiency of 87.59%, with a mean size of 152.37 ± 1.88 nm and zeta potential of −22.17 ± 0.69 mV. It released less than 7% of Scap in PBS over 48 h but up to 75% in mouse caecal content medium, and delayed small-intestinal Cap absorption (53.1% at 4 h vs. >80% for free Cap).
  • SCXN increased Cap AUC(0-∞) and T1/2 to 2.31 and 2.73 times those of free Cap, respectively, and reduced plasma clearance to 43.18% of free Cap. It also maintained higher intratumoral 5-Fu at 8–24 h.
  • In CT26-luc tumor-bearing mice, SCXN achieved a tumor inhibition rate of 71.78%, compared with 5.29% for Scap and 8.24% for Cap, and prolonged median survival to 33.5 d versus 14–15 d for saline, xylan, and Sxy groups. In MC38 tumors, SCXN achieved a TIR of 87.33%, with tumor volumes controlled below 500 mm³ versus 1547 mm³ for saline and 1111 mm³ for free Cap.
  • SCXN increased matured dendritic cells by 1.44× versus xylan+Cap and 2.64× versus Cap, increased intratumoral CD8+ T cells by 2.13× versus Cap, reduced Tregs to 56.6% of saline, and increased the CD8+ T cell/Treg ratio to 3.60× and 2.30× those of Cap and xylan+Cap, respectively. Fecal SCFA content increased by at least 37%, and butyric acid was 3.53× and 3.27× that of saline and Cap groups; beneficial taxa including Bifidobacterium, Roseburia, Akkermansia, and Faecalibaculum increased, while Desulfovibrio decreased.

Methods

  • The authors synthesized amphiphilic xylan–stearic acid (Sxy) and capecitabine–stearic acid (Scap) conjugates, prepared SCXN by film dispersion, and characterized it using NMR, MALDI-TOF MS, TEM, DLS, and HPLC.
  • In vitro release was tested in PBS, artificial gastric juice, and mouse caecal content medium. In vivo studies used CT26, CT26-luc, and MC38 mouse tumor models for biodistribution, pharmacokinetics, biocompatibility, anti-tumor efficacy, survival, immune cell analysis, 16S rDNA sequencing, and GC-MS SCFA measurement.
  • Mice were orally treated with 50 mg/kg Cap and 500 mg/kg xylan daily for three weeks. Statistical analyses included unpaired two-sided t-tests, one-way ANOVA with Tukey’s test, and log-rank tests, with n = 3–9 mice per group depending on the experiment.

Limitations

  • The paper does not include a dedicated limitations section. It states that no statistical method was used to predetermine sample size and that investigators were not blinded to allocation during experiments and outcome assessment.
  • Evident from the text: all efficacy and safety data come from mouse models (CT26, CT26-luc, MC38), sample sizes are small (often n = 3–9), and survival evaluation ended at Day 45. The anti-tumor dose of 50 mg/kg/day is less than a tenth of the clinical 61 mg/kg/day, so the free Cap comparison was at a low dose.
  • Evident from the text: microbiota and SCFA analyses are limited to fecal samples and 16S rDNA/GC-MS measurements, and the causal link between specific microbial taxa and tumor control is not fully established.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/s41467-023-40439-y

Cites

56 of 56 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Figures

Fig. 1 Construction and characterization of the Scap-loaded Sxy nanoparticle (SCXN). a Schematic illustration of combining chemotherapy and gut microbiota regulation for CRC treatment by SCXN. b Transmission electron microscopy images of the blank Sxy nanoparticle (BXN) and SCXN. Scale bars: 200 nm. For each group, 3 independent samples were tested, and 1 representative image is shown. c Size distribution of BXN and SCXN. d Zeta potentials of BXN and SCXN. e Drug release curves of free Scap and SCXN in phosphate buffered saline (PBS; pH 7.4) with or without mouse caecal content (MCC). f Variation of particle sizes of SCXN in PBS at pH 7.4 and artificial gastric juice (AGJ) from 0 to 8 h. g Cap absorption curves in the small intestine after the oral administration of free Cap or SCXN in mice. h Distribution of the 1-pyrenebutyric acid (PBA)-labelled Scap (Scap-PBA)-loaded Sxy nanoparticle (SCXN-PBA) and free Scap-PBA along the villi of jejunum at different time points after oral administration. Scale bar: 500 μm. Data represent the mean ± SD. For each group, n = 3 independent samples in (c–g), and n = 3 mice in h. Source data are provided as a Source Data file.
Fig. 1 Construction and characterization of the Scap-loaded Sxy nanoparticle (SCXN). a Schematic illustration of combining chemotherapy and gut microbiota regulation for CRC treatment by SCXN. b Transmission electron microscopy images of the blank Sxy nanoparticle (BXN) and SCXN. Scale bars: 200 nm. For each group, 3 independent samples were tested, and 1 representative image is shown. c Size distribution of BXN and SCXN. d Zeta potentials of BXN and SCXN. e Drug release curves of free Scap and SCXN in phosphate buffered saline (PBS; pH 7.4) with or without mouse caecal content (MCC). f Variation of particle sizes of SCXN in PBS at pH 7.4 and artificial gastric juice (AGJ) from 0 to 8 h. g Cap absorption curves in the small intestine after the oral administration of free Cap or SCXN in mice. h Distribution of the 1-pyrenebutyric acid (PBA)-labelled Scap (Scap-PBA)-loaded Sxy nanoparticle (SCXN-PBA) and free Scap-PBA along the villi of jejunum at different time points after oral administration. Scale bar: 500 μm. Data represent the mean ± SD. For each group, n = 3 independent samples in (c–g), and n = 3 mice in h. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 2 Biodistribution and pharmacokinetics of SCXN. a In vivo bioluminescence images of tumors (left) and the fluorescence of 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindotricarbocyanine iodide (DiR) (right) in CT26-luc tumor-bearing mice at 1, 4, 8, and 24 h after the oral administrated of free DiR or DiR-loaded Sxy nanoparticle (Sxy-DiR). Six biological replicates are shown. b Ex vivo fluorescent images of tissues from CT26 tumor-bearing mice at 1, 4, 8, and 24 h after orally administrated with free DiR or Sxy-DiR. c Ex vivo fluorescent images of intestines from CT26 tumor-bearing mice at 1, 4, 8, and 24 h after the oral administration of free DiR or Sxy-DiR. d The content of Cap in different organs at 1, 4, 8, and 24 h after the oral administration of free Cap or SCXN in CT26 tumor-bearing mice. e Content of Cap and 5-Fu in tumors at 1, 4, 8, and 24 h after the oral administration of free Cap or SCXN in CT26 tumor-bearing mice. f Plasma concentration-time curves of Cap after the oral administration of free Cap or SCXN in mice. Data represent the mean ± SD (n = 3 mice). Statistical significance was calculated using unpaired two-sided t-test, with Welch’s correction when variances are not equal. Source data are provided as a Source Data file.
Fig. 2 Biodistribution and pharmacokinetics of SCXN. a In vivo bioluminescence images of tumors (left) and the fluorescence of 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindotricarbocyanine iodide (DiR) (right) in CT26-luc tumor-bearing mice at 1, 4, 8, and 24 h after the oral administrated of free DiR or DiR-loaded Sxy nanoparticle (Sxy-DiR). Six biological replicates are shown. b Ex vivo fluorescent images of tissues from CT26 tumor-bearing mice at 1, 4, 8, and 24 h after orally administrated with free DiR or Sxy-DiR. c Ex vivo fluorescent images of intestines from CT26 tumor-bearing mice at 1, 4, 8, and 24 h after the oral administration of free DiR or Sxy-DiR. d The content of Cap in different organs at 1, 4, 8, and 24 h after the oral administration of free Cap or SCXN in CT26 tumor-bearing mice. e Content of Cap and 5-Fu in tumors at 1, 4, 8, and 24 h after the oral administration of free Cap or SCXN in CT26 tumor-bearing mice. f Plasma concentration-time curves of Cap after the oral administration of free Cap or SCXN in mice. Data represent the mean ± SD (n = 3 mice). Statistical significance was calculated using unpaired two-sided t-test, with Welch’s correction when variances are not equal. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 3 Biocompatibility evaluation in healthy mice receiving multi-dose treatments with different formulations. a Mice body weight variation during the treatment period. b Images of the hematoxylin and eosin (H&E)-stained sections of main organs. Scale bar: 50, 100, or 200 μm. c Biochemical parameters in blood. ALT alanine aminotransferase, AST aspartate aminotransferase, BUN blood urea nitrogen, CREA creatinine. Data represent the mean ± SD (n = 3 mice). Source data are provided as a Source Data file.
Fig. 3 Biocompatibility evaluation in healthy mice receiving multi-dose treatments with different formulations. a Mice body weight variation during the treatment period. b Images of the hematoxylin and eosin (H&E)-stained sections of main organs. Scale bar: 50, 100, or 200 μm. c Biochemical parameters in blood. ALT alanine aminotransferase, AST aspartate aminotransferase, BUN blood urea nitrogen, CREA creatinine. Data represent the mean ± SD (n = 3 mice). Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 4 In vivo anti-tumor effects in CT26-luc tumor-bearing mice receiving multi-dose treatments of different formulations. a In vivo bioluminescence imaging using an in vivo imaging system (IVIS). Three representative mice of each group are shown. Six biological replicates are conducted and shown in Supplementary Fig. 3. Red line boxes denote the dead mice. b Variation in relative tumor volumes calculated according to the amount of photons during the therapy period. c Variation in body weights during the therapy period. d Deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) immunofluorescence examination of the CT26-luc tumor sections. Scale bar: 200 μm. e H&E-stained intestine sections from the CT26-luc tumor-bearing mice at the end of the therapy period. Red dot line boxes denote the tumor burdens. Scale bar: 2 cm. f Survival curves of mice within 45 d post the 1st administration. Data represent the mean ± SD. For each group, n = 6 mice in (a–c) and (f), and n = 3 mice in (d and e). A two-sided log-rank (Mantel–Cox) test was used for the statistical comparison of the survival study. Source data are provided as a Source Data file.
Fig. 4 In vivo anti-tumor effects in CT26-luc tumor-bearing mice receiving multi-dose treatments of different formulations. a In vivo bioluminescence imaging using an in vivo imaging system (IVIS). Three representative mice of each group are shown. Six biological replicates are conducted and shown in Supplementary Fig. 3. Red line boxes denote the dead mice. b Variation in relative tumor volumes calculated according to the amount of photons during the therapy period. c Variation in body weights during the therapy period. d Deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) immunofluorescence examination of the CT26-luc tumor sections. Scale bar: 200 μm. e H&E-stained intestine sections from the CT26-luc tumor-bearing mice at the end of the therapy period. Red dot line boxes denote the tumor burdens. Scale bar: 2 cm. f Survival curves of mice within 45 d post the 1st administration. Data represent the mean ± SD. For each group, n = 6 mice in (a–c) and (f), and n = 3 mice in (d and e). A two-sided log-rank (Mantel–Cox) test was used for the statistical comparison of the survival study. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 5 In vivo anti-tumor effects in MC38 tumor-bearing mice receiving multi-dose treatments of different formulations. a, b Tumor images (a) and average tumor weights (b) on day 21. c Variation in tumor volumes during the therapy period. d TUNEL immunofluorescence examination of MC38 tumor sections. Scale bar: 200 μm. Data represent the mean ± SD. For each group, n = 6 mice in (a–c), and n = 3 mice in (d). Statistical significance was calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.
Fig. 5 In vivo anti-tumor effects in MC38 tumor-bearing mice receiving multi-dose treatments of different formulations. a, b Tumor images (a) and average tumor weights (b) on day 21. c Variation in tumor volumes during the therapy period. d TUNEL immunofluorescence examination of MC38 tumor sections. Scale bar: 200 μm. Data represent the mean ± SD. For each group, n = 6 mice in (a–c), and n = 3 mice in (d). Statistical significance was calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 6 SCXN promotes the anti-tumor immune responses. Analysis of the numbers of immune cells in CT-26 tumor-bearing mice treated with multi-doses of different formulations via flow cytometry or immunofluorescence assay. a, b Numbers (a) and percentages (b) of mature dendritic cells (DCs; CD80+CD86+ cells gated on CD11c+ cells) in draining lymph nodes. c Numbers of the CD8+ T cells per mg of tumor. d Percentage of CD8+ T cells in the total CD3+ cell population in tumors. e Immunofluorescence images of tumor sections to examine the CD8+ T cells (red fluorescence) infiltration. Green fluorescence: CD31. Scale bar: 200 μm. f, g Numbers (f) and percentage (g) of regulatory T cells (Tregs, CD4+Foxp3+ cells gated on CD3+CD4+ cells) in tumors. h Ratio of CD8+ T cells to Tregs in tumors. Data represent the mean ± SD (n = 3 mice). Statistical significance was calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.
Fig. 6 SCXN promotes the anti-tumor immune responses. Analysis of the numbers of immune cells in CT-26 tumor-bearing mice treated with multi-doses of different formulations via flow cytometry or immunofluorescence assay. a, b Numbers (a) and percentages (b) of mature dendritic cells (DCs; CD80+CD86+ cells gated on CD11c+ cells) in draining lymph nodes. c Numbers of the CD8+ T cells per mg of tumor. d Percentage of CD8+ T cells in the total CD3+ cell population in tumors. e Immunofluorescence images of tumor sections to examine the CD8+ T cells (red fluorescence) infiltration. Green fluorescence: CD31. Scale bar: 200 μm. f, g Numbers (f) and percentage (g) of regulatory T cells (Tregs, CD4+Foxp3+ cells gated on CD3+CD4+ cells) in tumors. h Ratio of CD8+ T cells to Tregs in tumors. Data represent the mean ± SD (n = 3 mice). Statistical significance was calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 7 Regulating gut microbiota by SCXN. CT-26 tumor-bearing mice were treated with different formulations for 20 d and fecal samples were collected for 16 S rDNA sequencing before (Day 0), in the middle of (Day 10), and at the end of (Day 20) the treatment. a Microbial α-diversity in terms of Shannon index at the ASV level. b Microbial β-diversity NMDS analysis based on Bray-Curtis distance at the ASV level at the end of the treatment. c Heatmap showing relative abundance of the gut microbiota at the family level (displayed as normalized Z-score). d Barplot showing relative abundance of the gut microbiota at the genus level. e Change of the relative abundance of Akkermansia, Roseburia, Fecalibaculum, and Bifidobacterium during 20-day treatment process. f LefSe analysis cladogram representing the significantly different taxas between different groups from phylum to genus levels at the end of the treatment (LDA > 2, p < 0.05). g Summary of fecal SCFA levels and butyric levels in feces from CT26 tumor-bearing mice after different treatments. Data represent the mean ± SD (n = 5 mice). Statistical significance was calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.
Fig. 7 Regulating gut microbiota by SCXN. CT-26 tumor-bearing mice were treated with different formulations for 20 d and fecal samples were collected for 16 S rDNA sequencing before (Day 0), in the middle of (Day 10), and at the end of (Day 20) the treatment. a Microbial α-diversity in terms of Shannon index at the ASV level. b Microbial β-diversity NMDS analysis based on Bray-Curtis distance at the ASV level at the end of the treatment. c Heatmap showing relative abundance of the gut microbiota at the family level (displayed as normalized Z-score). d Barplot showing relative abundance of the gut microbiota at the genus level. e Change of the relative abundance of Akkermansia, Roseburia, Fecalibaculum, and Bifidobacterium during 20-day treatment process. f LefSe analysis cladogram representing the significantly different taxas between different groups from phylum to genus levels at the end of the treatment (LDA > 2, p < 0.05). g Summary of fecal SCFA levels and butyric levels in feces from CT26 tumor-bearing mice after different treatments. Data represent the mean ± SD (n = 5 mice). Statistical significance was calculated using one-way ANOVA with Tukey’s multiple comparisons test. Source data are provided as a Source Data file.CC BY · PubMed Central

Verification

Claims the paper makes

76 Own result · 26 Own interpretation · 14 Not checked yet · 10 No full text of the sources · 9 Own method · 9 No basis given · 6 Partly supported · 1 Supported so far · accuracy not yet measured

Verify all 22 references — free account

Show all 77 claims

  • Probably not a substantial claim. 23
  • It may state two separate claims as one. 55
  • The quote only partly states this claim. 1
74 of 74
  • The paper relies on this without support.

    Prebiotic delivery into the gut is a more controllable method for gut microbiota modulatory therapy than traditional intervention.

    “Compared with traditional intervention, prebiotics delivery into the gut is a more controllable method for gut microbiota modulatory therapy.”
  • The paper relies on this without support.

    Capecitabine lacks a carrier that can prolong its half-life.

    “Capecitabine (Cap), the first-line chemotherapeutic agent for CRC, lacks a carrier that can prolong its half-life.”
  • The paper relies on this without support.

    Large-scale application of FMT is limited by the lack of management standards and risk of serious infection.

    “However, large-scale application of FMT is limited by the lack of management standards and risk of serious infection.”
  • The paper relies on this without support.

    The specific prebiotic role of xylan in treating cancer remains unknown.

    “The specific prebiotic role of xylan in treating cancer remains unknown.”
  • The paper relies on this without support.

    The negative charge of SCXN is beneficial to avoid clearance by the monocyte-phagocyte system.

    “This charge was beneficial to avoid the clearance by the monocyte-phagocyte system.”
  • The paper relies on this without support.

    The mouse CRC models used in this study are predictive of human CRC therapy outcomes.

    “In conclusion, we demonstrated the potential of the combination of gut microbiota modulation and chemotherapy in CRC therapy via a single drug delivery system using prebiotics.”
  • The paper relies on this without support.

    The gut microbiota of mice is representative of human gut microbiota for the purpose of xylan metabolism and CRC therapy.

    “The reason might be that the SCFA-producing probiotics, which can degrade xylan, represented by Lactobacillus and Bifidobacterium56, had a much lower abundance in the intestines of colon tumor-bearing mice than that in the normal ones.”
7 of 7

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

1 of 1

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Metadata

Authors
Lang, T., Zhu, R., Zhu, X., Yan, W., Li, Y., Zhai, Y., Wu, T., Huang, X., Yin, Q., & Li, Y.
Affiliations
  • Chinese Academy of Sciences (CN)
  • Nanjing University of Chinese Medicine (CN)
  • Nanjing Medical University (CN)
Corresponding author
  • Li, Yu
  • Yin, Qi
  • Li, Yaping
Venue
Nature Communications
Published
2023-08-07
Volume
14
Issue
1
Pages
4746-4746
ISSN
2041-1723 (ISSN-L)
OpenAlex
W4385638731
Cited by
133
References
56
Citations by year
2026: 47 · 2025: 48 · 2024: 37 · 2023: 1
FWCI
7.62
Type
journal article
Language
en
Open access
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (31930066, 32130058, 32171315)
  • Natural Science Foundation of Shandong Province (ZR2019ZD25)
Topics
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advanced Drug Delivery Systems
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
Keywords
colorectal cancer, capecitabine, gut microbiota modulation, oral drug delivery, drug half-life extension, short-chain fatty acids, antitumor immunity
MeSH terms
Gastrointestinal Microbiome, Capecitabine, Animals, Colorectal Neoplasms, Mice, Nanoparticles, Prebiotics

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