Protocol detail

RPPA-Based PI3K Pathway Characterization in TNBC PDX Models

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RPPA was performed as previously described 29 (link) for 48 of the 57 TNBC PDX (9 PDX were established after the RPPA analysis). We calculated a PI3K pathway score with normalized data to assess pathway activation. Scores were obtained by calculating the sum for positive protein components (PI3K p110 subunit β, p-AKT1 (Ser473), p-AKT1 (Thr308), p-4E-BP1, p-p70-S6 kinase, p-S6 ribosomal protein; Cell Signaling Technology®) and subtracting the negative components of the pathway (PTEN, Cell Signaling Technology®). Eleven PDX classified as unstable (UNS) in Lehmann's classification were excluded from the analysis shown in Figure 3B.

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Coussy F., Lavigne M., de Koning L., Botty R.E., Nemati F., Naguez A., Bataillon G., Ouine B., Dahmani A., Montaudon E., Painsec P., Chateau-Joubert S., Laetitia F., Larcher T., Vacher S., Chemlali W., Briaux A., Melaabi S., Salomon A.V., Guinebretiere J.M., Bieche I, & Marangoni E. (2020). Response to mTOR and PI3K inhibitors in enzalutamide-resistant luminal androgen receptor triple-negative breast cancer patient-derived xenografts. Theranostics, 10(4), 1531-1543.

Publication 2020

p-AKT1 (Ser473 p-AKT1 (Thr308), p-4E-BP1 p-p70-S6 kinase p-S6 ribosomal protein

4e bp1 Akt1 P70 s6 kinase Pi3k Protein Pten Ribosomal protein Subunit

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Variable analysis

independent variables

RPPA analysis

dependent variables

PI3K pathway score
Pathway activation

control variables

48 out of 57 TNBC PDX (9 PDX were established after the RPPA analysis)
Eleven PDX classified as unstable (UNS) in Lehmann's classification were excluded from the analysis shown in Figure 3B

positive controls

PI3K p110 subunit β
P-AKT1 (Ser473)
P-AKT1 (Thr308)
P-4E-BP1
P-p70-S6 kinase
P-S6 ribosomal protein

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