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. Author manuscript; available in PMC: 2023 Jan 1.
Published in final edited form as: Adv Mater. 2021 Nov 14;34(2):e2100096. doi: 10.1002/adma.202100096

Figure 1.

Figure 1.

Proteomic, transcriptomic, and spatial omic analysis of patient tumors and establishment of CRPC-Adeno Matrigel organoids. A) Mass spectrometry analysis of ECM components (left) and integrin signaling-related components (right) in primary CRPC-Adeno tumors relative to adjacent normal tissue (average of n = 3). B) RNA-seq transcriptomic analysis of ECM components and integrins in CRPC-Adeno and CRPC-NEPC patient tumor biopsies, as compared to benign tissues (n = 74 CRPC-Adeno, n = 37 CRPC-NEPC, n = 31 benign samples). Data presented as mean ± SEM. C) Multiplexed single-cell spatial omics RNA analysis of prostate tumors. Left column: Immunostaining of EZH2 on CRPC-NEPC prostate tumors. Zoomed in images of single-cell EZH2 distributions (green) in the second and third rows. Right column: FISH based detection of single COL1A1 and β-actin RNA molecules in the same area of the CRPC-NEPC prostate tumors using a FISH signal amplification assay, HCR. Zoomed-in images of single-cell RNA distributions of COL1A1 (yellow) and β-actin (magenta) in the second and third rows. Data representative of two patient biopsies. D) SHG microscopy images of collagen fibers in CRPC-NEPC tissues. The tissue samples were embedded in paraffin and stained with Synaptophysin (SYP). The SHG of tissue samples were imaged on a deparaffinized unstained slide using a two-photon microscope. Data representative of two patient biopsies. E) IHC staining of CRPC-NEPC and CRPC-Adeno patient biopsies. SYP and NKX3.1 staining for CRPC-NEPC and CRPC-Adeno, respectively, and expression of integrin α2 and β1. Data representative of two patient biopsies. F) Development and characterization of a new CRPC-Adeno organoid (OWCM-1358) from a patient tumor biopsy. Nanostring analysis of hallmark gene expression in multiple metastatic sites and derived Matrigel organoids. G) H&E and IHC staining of CRPC-Adeno patient and OWCM-1358 Matrigel organoid that demonstrated CRPC-Adeno gene signature. H) Nanostring-based comparative analysis of prostate cancer-associated genes among CRPC-Adeno (OWCM-1358) and CRPC-NEPC Matrigel organoids (OWCM-154, OWCM-155). I) IHC staining of CRPC-NEPC and CRPC-Adeno Matrigel organoids. SYP and NKX3.1 staining for CRPC-NEPC and CRPC-Adeno, respectively, and expression of integrin α2 and β1. Data representative of two samples.