Skip to main content
. 2024 Sep 26;5(10):101748. doi: 10.1016/j.xcrm.2024.101748

Figure 5.

Figure 5

Genotyped-phenotyped CD PDOs can serve as platforms for personalized therapeutics

(A) Schematic outlines the strategy for therapeutic reversal of the disease driver phenotypes in each molecular subtype of CD (B–I) and for crossover efficacy across the subtypes (J–M).

(B) Mean fluorescent intensity of SA-βGal staining of PDOs by flow cytometry (B- yellow cluster CD PDOs separated into groups that responded (R) or not (NR) to anti-TNF-α biologics or were naive to that treatment). Data represents 2–4 technical repeats on 4 healthy and 6 CD PDOs. Statistical significance was assessed by one-way ANOVA.

(C) Histogram (left) and violin plots (right) show the % changes in the median fluorescence intensity when CD PDOs were treated with senotherapeutics (2.5 μM PAC, pacritinib; 1 mM Met, metformin). Statistical significance was assessed by one-way ANOVA.

(D–E) Inverted images displayed (D) are representative of ∼10 fields/sample of max-projected z stacks of CD PDOs stained with SPiDER-SA-β-Gal. Bar graphs (E) display the quantification of staining. Scale bar, 100 μm. Statistical significance was assessed by one-way ANOVA.

(F) Schematic outlines the 3 major steps in bacterial clearance assays, and the concomitant assessment of supernatants for cytokines by multiplexed ELISA.

(G) Heatmap displays the results of hierarchical agglomerative clustering of AIEC-LF82-challenged healthy and CD-EDMs using the cytokine profiles determined by mesoscale (MSD). See Table S7.

(H) Bar plots show the abundance of bacteria retained within healthy (H) and IDICD EDMs at 8 h after infection. Statistical significance was assessed by t test.

(I) Line plots show the pre-(UN) and post-treatment (Rx) effect of a balanced PPARα/g dual agonist (1 μM PAR5359) on the abundance of bacteria at the 8 h time point in healthy (left) and IDICD (right) EDMs. Statistical significance was assessed by t test.

(J and K) Violin plots show the % change in median fluorescence intensity with PAC (J) or PAR5359 (K), as determined on S2FCD PDOs with the highest senescence in (B). Statistical significance was assessed by t test.

(L and M) Line plots show the untreated and effect of PAR5359 (L) or PAC (M) treatment on the abundance of bacteria at the 8 h time point in IDICD EDMs with the highest bacterial load in H. Statistical significance was assessed by t test. See Table S2 for subjects analyzed in each assay.