Fig 2: The gut microbiota profiles in cholecystectomy patients and mice models.a α-diversity of the gut microbiota at all levels in HC and GBx patients (two-tailed Welch’s t test). b NMDS of bacterial communities based on Bray-Curtis similarity. c VIP score of the PLS-DA in gut microbiota. VIP scores were used to rank the ability of different taxa to discriminate between the two groups. A taxon with VIP score > 2.0 was considered important in the discrimination. d Index (α-diversity) of the gut microbiota between Sham+AOM/DSS and GBx+AOM/DSS mice. (two-tailed Welch’s t test, n = 7 vs 9). The center line of the boxplot represents the median; the box spans from the first quartile (Q1) to the third quartile (Q3); the whiskers extend from the box to the maximum and minimum values within 1.5 times the interquartile range (IQR) from the box. e NMDS of bacterial communities based on Bray-Curtis similarity (n = 7 vs 9). f VIP score of the PLS-DA. VIP scores were used to rank the ability of different taxa to discriminate between the two groups. A taxon with VIP score > 2.0 was considered important in the discrimination (n = 7 vs 9). g Relative abundance (percent reads) of B. breve between healthy and cholecystectomy patients based on metagenomics sequencing. h Relative abundance (percent reads) of R. gnavus between healthy and cholecystectomy patients based on metagenomics sequencing. i The relative abundance of B. breve between the Sham+AD and GBx+AD group (n = 7 vs 9). j Relative abundance of R. gnavus between Sham+AOM/DSS and GBx+AOM/DSS mice (n = 7 vs 8). k qPCR analysis of B. breve abundance in Sham and GBx mice (n = 5 vs 6). l qPCR analysis of R. gnavus in Sham and GBx mice (two-tailed t test, n = 5 vs 6). m qPCR analysis of B. breve abundance in APC-Sham and APC-GBx mice (n = 6/group). n qPCR analysis of R. gnavus in APCmin/+ mice (two-tailed Welch’s t test, n = 6/group). Data are shown as the mean ± SEM. P values were determined by two-tailed Mann-Whitney U test for g–k, m, and were indicated in each figure. HC healthy control, AD AOM/DSS, NMDS nonmetric multidimensional scaling, VIP variable importance in projection, B. breve, Bifidobacterium breve, R. gnavus Ruminococcus gnavus. Source data are provided as a Source Data file.

Fig 3: Cholinergic innervation of APCs is essential for yki3SA-tumor-induced Akh release and host wasting.a Cholinergic boutons indicated by Cha > Syt-GFP (green) in the somas of adult APCs (red, anti-Akh) at day 4. b Trans-tango system indicating overlaps between cholinergic innervating neurons (GFP) and APCs (red, anti-Akh). c Whole body Akh and tobi expression (left, n = 3, 5 flies/replicate) and metabolic changes such as TAG and TRE storages (right, n = 4, 5 flies/replicate) of adult flies with activation of cholinergic neurons induced at 29 °C for 12 h. d Upstream cholinergic boutons indicated by Syt-GFP (left, green) in APCs (left, red, anti-Akh) and quantification of bouton numbers (right, n = 3) of yki3SA-tumor-bearing flies (LexA + GAL4) at day 6. e‒h Bloating phenotype (e, up), gut tumors (e, middle, green) and quantification (f, n = 3), image of APCs (e, bottom, red, anti-Akh), quantification of APC masses (area) and intracellular Akh amounts (g, n = 3), whole body Akh and tobi expression (h, n = 3, 5 flies/replicate), abdomen bloating rates (i, left, n = 4) and TAG and TRE storages (i, right, n = 4, 5 flies/replicate) of yki3SA-tumor-bearing flies with synapse disruption in cholinergic neurons at day 6 (LexA + GAL4). j The schematic model illustrating the release of Akh and its regulation by tumor-secreted Pvf1 and ECM-associated neural innervation in yki3SA-tumor-bearing flies, leading to organ wasting. Data are presented as mean ± SEM. Each dot represents one biological replicate. Statistical analysis was conducted by two-tailed unpaired t-test (c, d), one-way ANOVA with Bonferroni’s multiple-comparisons test (f‒i). *P < 0.05.

Fig 4: Oral administration of D5P attenuates CRC development in APC mutant mice. A Kaplan–Meier survival analysis of colon adenocarcinoma patients stratified by PNP expression. Data from GEPIA database. B Rectal prolapse observation showing that the onset of CRC in APCmin/+ mice treated with D5P was delayed. C Survival probability analysis of APCmin/+ mice treated with or without D5P (4 mM) in drinking water. D Representative colon images of the APCmin/+ mice treated with or without D5P (4 mM). E The quantification of tumor number presented in (D). Colon tumor number of APCmin/+ mice treated with or without D5P (4 mM). F Representative H&E staining of colon tumor from mice treated with or without D5P (4 mM). G Large field view of YAP staining in colon sections of mice treated with or without D5P (4 mM). The numeric labeled regions were zoom in and showed in (H) and (I). H A representative image of non-cancerous area in (G). YAP staining of the non-cancerous area of colon from mice treated with or without D5P (4 mM). I A representative image of cancerous area in (G) and quantitative results of YAP staining in the cancerous area of colon from mice treated with or without D5P (4 mM). J Schematic diagram showing the anti-cancer function of D5P via the MYH9/LATS1-YAP-PNP-D5P negative feedback loop. *, P < 0.05; **, P < 0.01

Fig 5: Akh is essential for tumor-induced host wasting in Drosophila.a‒h Representative images of abdomen bloating (a, up), Akh production in APCs (a, middle, red, anti-Akh), and gut tumors (a, bottom, GFP), quantification of APC masses (area) (b, left, n = 3) and intracellular Akh amounts (b, right, n = 3), circulating Akh levels in the hemolymph (c, left, dot-blot; right, quantification, n = 4), quantification of tumor mass (GFP intensity/gut area) (d, n = 3), whole body Akh, AkhR, and tobi mRNA levels (e, n = 4‒6, 5 flies/replicate), bloating rates (f, n = 3, 20 flies/replicate), metabolic dysregulation such as TAG and TRE storages (g, n = 4, 5 flies/replicate), circulating TRE levels in the hemolymph (h, n = 4, 40 flies/replicate) and food intakes (i, n = 4, 20 flies/replicate) of adult yki3SA-tumor-bearing flies with or without Akh∆ mutation (AkhSAP/AkhA) at day 8. j Heatmap indicating both yki3SA-tumor- and Akh-dependent differentially expressed genes in the whole-body. Data are presented as mean ± SEM. Each dot represents one biological replicate. Statistical analysis was conducted by two-tailed unpaired t-test (b, c, e, left) and one-way ANOVA with Bonferroni’s multiple-comparisons test (d, e, right, f‒i). *P < 0.05.