Journal: Cancers

Article Title: Drug-Induced Resistance and Phenotypic Switch in Triple-Negative Breast Cancer Can Be Controlled via Resolution and Targeting of Individualized Signaling Signatures

doi: 10.3390/cancers13195009

Figure Lengend Snippet: TNBC tissues are represented by different patient-specific signaling signatures, majority of which do not include EGFR. ( A ) Fold changes in expression levels of EGFR and pEGFR in TNBC and non-TNBC tumors are shown. Y1068 and Y1173 are EGFR phosphorylation sites; ( B ) Examples for patient-specific sets of active unbalanced processes are shown. Each sample harbors a set of 1–3 active unbalanced processes (PaSSS), represented schematically by a barcode. In each barcode active unbalanced processes are represented by black or gray squares, inactive white. Negative/positive amplitude denotes how the patients are correlated with respect to a particular process. Suggested PaSSS-based therapies appear below each barcode; ( C ) Heterogeneity index of 3 subgroups, represented by a ratio between the number of distinct PaSSSs and the number of samples in each subset, is shown for the TNBC subset of tissues, the entire set (3467 samples from 11 cancer types) and the subset of non-TNBC samples. (Abbreviations: TNBC—Triple Negative Breast Cancer, PaSSS—Patient-specific signaling signature, EGFR—Epidermal Growth Factor Receptor, VEGFR2—Vascular Endothelial Growth Factor Receptor 2, Her2—Human Epidermal growth factor Receptor 2, Src—Proto-oncogene tyrosine-protein kinase Src).

Article Snippet: The following conjugated antibodies were used: anti-p-EGFR (Y1068) (R&D Systems, Minneapolis, MN, USA, cat. no. IC3570G), anti-p-ERK2 (Thr202/Tyr204) (BioLegend, San Diego, CA, USA, cat. No. 675503), anti-p-S6 (Ser235/236) (BioLegend, cat. no. 608605), and anti-GAPDH (Santa Cruz Biotechnology, Dallas, Texas, USA, cat. no. sc-47724AF594).

Techniques: Expressing, Phospho-proteomics

Journal: JCI Insight

Article Title: Interim report on engineered NK cell trial in lung cancer refractory to immune checkpoint inhibitors

doi: 10.1172/jci.insight.186890

Figure Lengend Snippet: Utilizing the multicolor immunofluorescence histochemical staining, we examined lung tumor tissue from patient 006 on 3 occasions using DAPI (blue) for nuclear staining, pan-CK (gray) for lung parenchyma, CD45 (purple) for lymphocytes, CD57 (cyan) and CD56 (green) for NK cells, CD3 (yellow) for T cells, and EGFR (red) combined with CD56 or CD57 to identify NK cells transduced with the tEGFR-sIL15 construct. ( A ) FFPE tissue from a pretreatment tumor biopsy from patient 006 shows the presence of lung parenchyma (DAPI + pan-CK + ) but the absence of tEGFR + (red) transduced CD57 + or CD56 + NK cells. ( B ) FFPE tissue from the skinny needle tumor biopsy collected 24 hours following the fourth and final infusion of sIL15_TRACK NK cells without observable of tEGFR + (red) transduced CD57 + (cyan) or CD56 + (green) cells. ( C ) FFPE tissue from an excisional tumor biopsy collected 7 days following the fourth and final infusion of sIL15_TRACK NK cells shows sIL15_TRACK NK cells identified in DAPI + CD45 + cells by coexpression of tEGFR (red) with CD57 (cyan), as marked by white arrows. ( D ) Selected individual markers of the boxed region shown in C are shown. Scale bars: 10 μm ( A – C ); 10 μm ( D ).

Article Snippet: For assessment of tumor infiltration of NK cells, primary antibodies were as follows: EGFR (clone 31G7, Novus), CD3 (clone LN10, Leica), CD56 (clone MRQ-42, Cell Marque), CD45 (clone D9M81, Cell Signaling Technologies), CD57 (clone HNK-1, Biolegend), and pan-cytokeratin (clone AE1/AE3, Dako).

Techniques: Immunofluorescence, Staining, Transduction, Construct