Figure 5.

3D imaging reveals two distinct DCIS growth patterns with strong correlation to invasive progression

(A) Examples of 3D whole-mount imaging of DCIS lesions in PDX models, showing replacement growth (left panel) and expansive growth (right panel). Cells of human origin are marked with Ku80 (green), myoepithelial cells are marked with αSMA (magenta).

(B) Volumes of DCIS lesions for replacement and expansive growth. Data are represented as mean ± SEM. Two-tailed Mann-Whitney test. ∗∗p < 0.01.

(C) Width to length ratios of DCIS lesions for replacement and expansive growth, revealing a more elongated shape for lesions with replacement growth vs. a more spherical shape for lesions with expansive growth. Data are represented as mean ± SEM. Two tailed Mann Whitney test. ∗∗p < 0.01.

(D) Invasive progression rates of DCIS samples showing replacement growth, mixed growth, and expansive growth, respectively.

(E–G) 3D morphologies of FFPE blocks from patients with DCIS. FFPE blocks were tissue-cleared and imaged for αSMA (cyan), nuclei (gray), and second harmonics generation detecting collagen 1 (red).

(E) Example of globular DCIS morphology, representing expansive growth. Left, 3D visualization of the intact FFPE block. Middle, optical section through the indicated area in the 3D view. Right, 3D surface reconstruction of DCIS ducts in the indicated area.

(F) Example for tubular DCIS morphology, representing replacement growth. Left, 3D view of the intact FFPE block. Middle, optical section through the indicated area. Right, 3D surface reconstruction of the indicated area.

(G) Scheme illustrating DCIS morphologies in human resections that can be distinguished as globular expansion and tubular growth along ducts.

(H) Volcano plot of differentially expressed genes in DCIS lesions for replacement vs. expansive growth.

(I) Immunofluorescence staining of ECM1 (white) in a DCIS lesion showing replacement growth (top panels), a lesion showing expansive growth (middle panels), and a normal mammary duct (bottom panels). High extracellular expression of ECM1 protein is detected in the expansive DCIS lesion. Cells of human origin are marked with Ku80 (green), myoepithelial cells are marked with αSMA (magenta).

(J) Quantification of extracellular ECM1 expression in DCIS lesions for expansive or replacement growth and normal mammary ducts. Lines indicate Q¹, median, and Q³. Two-tailed Mann-Whitney test. ∗∗∗p < 0.001, ∗p < 0.05, ns, not significant. See also Figure S5.