Fig. 1

Overview of study design. A Formalin-fixed, paraffin-embedded (FFPE) primary and/or metastatic tumor surgical biopsy specimens were prospectively obtained from patients with PDAC, breast cancer, and other solid tumor malignancies as part of an IRB-approved Molecular Tumor Board (MTB) study at the Inova Schar Cancer Institute (ISCI). B Workflow diagram depicting tissue specimen analysis. Tumor epithelium (5–10 µm²) from tissue specimens was harvested via LMD at a ratio of 1 mm² LMD area per 2.5 µl buffer into a lysis buffer containing 50 mM Bond-Breaker TCEP Solution, 225 mM Tris–HCl, 4% v/v sodium dodecyl sulfate (SDS), 10% v/v glycerol, in MilliQ Type I water. RPPA analysis of the LMD enriched tissue lysates was performed by Theralink Technologies, Inc., as previously described [32, 33]. C Representative micrographs of tissue thin sections stained with hematoxylin and eosin (H&E) mounted onto a glass slide histopathological assessment (left, 5 µm), and onto a slide containing a polyethylene naphthalate (PEN) membrane after LMD harvest of tumor epithelium (right, 8 µm). Clinical NGS was performed by a commercial sequencing laboratory