TEST:

OncoSignal™

Conclusion The OncoSIGNal test provides a robust and quantitative method to characterize low ER and HER2 1+ IHC breast cancer samples. In addition to detecting and quantifying the relevant cell signaling pathways, the mRNA markers can be used to elucidate the results of low ER and HER2 1+ IHC staining, enabling the identification of patients who may benefit from targeted therapy.

The findings from this window-of-opportunity study highlight the need for larger cohort analyses to validate these initial observations and further investigate the clinical impact of these racial differences in breast cancer biology and treatment response.

We show that we are able to quantitatively measure the capacity of drugs to inhibit activity of the PI3K pathway and correlate its activity directly to biological response. Pathway activity measurement is more predictive for the cell growth inhibition than measurement of KI-67 mRNA expression. Clinical studies are in progress to evaluate the value of measuring pathway activity as predictive biomarker for response to PI3K, AKT or mTOR inhibitors in patients.

Higher Ki67 gene expression was associated with recurrences (p=0.042) Increased PI3K pathway activity, independent of PIK3CA mutations, may play a role in the recurrence of early-stage breast cancer with low and intermediate 21-gene RS. Pathway analysis can help to identify high-risk patients in this setting.

"The InnoSIGN team...during the San Antonio Breast Cancer Symposium (SABCS), from Wednesday December 6 to Friday 8 December, 2023...An abstract by NYU-Langone and two abstracts by InnoSIGN have been selected for poster presentation in the scientific program."

"The InnoSIGN team is excited to meet you at this year’s AMP Annual Meeting & Expo in Salt Lake City, US, from Tuesday November 14 to Saturday 18 November, 2023...Applications of OncoSIGNal pathway profiling platform in breast cancer and bladder cancer will be presented at the InnoSIGN corporate workshop..."

"...InnoSIGN Inc...announces the offering of its OncoSIGNal pathway profiling platform for clinical use as a Laboratory Developed Test (LDT) through InnoSIGN’s high-complexity CLIA laboratory...OncoSIGNal is the first platform identifying the tumor-driving cell signaling pathways in tumor tissue."

Based on the two week Ki67-IHC, patients were randomized (Ki67 ≥1%) to receive either Letrozole + Ribociclib or standard chemotherapy until surgery, or continued (Ki67 < 1%) Letrozole mono therapy. Positive ER staining does not always relate to a high ER pathway activity, possibly explaining that not all patients respond equally well to ER inhibition therapy. Activation of hormonal pathways (ER, but also AR) appear to be predictive for early (2 weeks) response towards Letrozole as assessed by Ki67 staining. Involvement of non-hormonal pathways is associated with less effective response towards Letrozole alone.

Compared to ER positive primary tumors we observed in metastatic ER positive tumors less frequent activation of the ER and more frequent activation of the PI3K pathway, suggesting that the latter pathway is associated with a more aggressive tumor phenotype. The triple negative subtype is characterized predominantly by activation of the PI3K, MAPK, AR and/or HH pathways, which may create new options for personalized targeted therapy of this subtype. OncoSIGNal can be used to determine the tumor driving signaling pathways in breast cancer patients, guiding selection of personalized targeted therapies.

Our observation suggests that distinct biomarkers may be differently suited to predict response to PI3K inhibitors in TNBC. This may include expanding the number of patients for which PI3K/Akt pathway inhibition might be a therapeutic option, as well as explaining the lack of response in PI3K mutated tumors.

Our data suggest that RNA expression-based pathway activity and DNA mutational and methylation analyses provide complementary information about the molecular landscape of TNBC. PI3K mutations are early drivers and lead to distinct DNA methylation signatures; however, they are not associated with increased PI3K pathway activity by RNA expression. In contrast, TNBC without PI3K mutations may show activation of the PI3K pathway in the absence of PI3K/Akt mutations.

The OncoSignal seven-pathway LDT test (ER, AR, PI3K, MAPK, Hedgehog, TGF- β, and Notch) is a robust and reliable method to quantify signaling pathway activity. By comparing with reference ranges from non-malignant tissues, the test can be used to identify tumor driving pathways. This opens new options for selection of targeted therapies for hard-to-treat breast cancer patients.

Our data suggest that RNA expression-based pathway activity and DNA mutational and methylation analyses provide complementary information about the molecular landscape of TNBC. PI3K mutations are early drivers and lead to distinct DNA methylation signatures; however, they are not associated with increased PI3K pathway activity by RNA expression. In contrast, TNBC without PI3K mutations may show activation of the PI3K pathway in the absence of PI3K/Akt mutations.

"InnoSIGN B.V. and the Catharina Hospital Eindhoven receive a grant of almost € 700 000 by the European subsidy program OPZuid for the 'STA-OP' multicenter prospective clinical trial, starting April 2023 (for 3 years). The objective of this trial is to identify new personalized targeted therapies based on 'Signal Transduction pathway Activity for recurrent Ovarian cancer Patients'...In this clinical trial InnoSIGN’s proprietary OncoSIGNal technology will be used to identify the tumor driving cell signaling pathways in the patient tumor tissue to direct patients to personalized therapies with targeted drugs that inhibit the activity of these pathways."

F.C. de Jong (Erasmus Medical Center, NL) for response prediction to BCG in bladder cancer.

Conclusions Using the OncoSIGNal test, actionable STP profiles were determined in samples from melanoma and BCC patients, creating opportunity for personalized targeted treatment. The OncoSIGNal test will be employed in the FINPROVE trial to identify patient specific actionable targets in several tumor types using tissue specific references.

"...Institut Gustave Roussy, published in Clinical Cancer Research their findings using OncoSIGNal for prostate cancer...'Genomic profiling of metastatic castration-resistant prostate cancer samples resistant to androgen-receptor pathway inhibitors'...It is shown that Low AR activity, activation of stemness programs, and Hedgehog pathway were associated with primary androgen receptor axis inhibitors’ resistance, whereas most acquired resistance was associated with subclonal evolution, AR-related events, and neuroendocrine differentiation."

"Joy Zhou Done from Johns Hopkins Hospital will present her findings on racial differences in ER staining positive breast cancer during the American Society of Breast Surgeons 24th Annual Meeting in Boston, US...By using OncoSIGNal there were measurable differences in ER pathway activity between White and Black patients prior to hormonal treatment, although all tumors were ER+ by immunohistochemistry."

"InnoSIGN presents the use of OncoSIGNal in triple breast cancer patients to identify new targeted treatment options."

By using the OncoSIGNal test high aberrant STP activities could be determined in TNBC patient samples. Results show that different pathways are aberrantly active, depending on the patient sample, which opens the opportunity for personalized targeted treatment of individual TNBC patients.Next steps: The TNBC data set will be extended to further profile the STP activity in relation to different clinical outcomes and to investigate new opportunities for targeted therapies.

A biopsy of a metastatic site was obtained prior start of ICI therapy (nivolumab or pembrolizumab). Signal transduction pathway activity of metastatic samples, taken prior start of ICI therapy, from patients with advanced melanoma, suggest that TGF-β did not correlate with response nor PFS but increased HH and MAPK STP activity may relate towards a worse PFS from ICI therapy.

By combining multiple types of molecular analyses, it was uncovered that this patient was eligible for novel clinical trials containing MEK and PD-L1 checkpoint inhibitors. Digital tools for genomic interpretation, clinical trials matching, and molecular tumor board discussions not only provided clinical decision support but also accelerated the path to treatment options.

Therefore, RL as an alternative for NAC merits further investigation in follow-up studies. ClinicalTrials.gov: NCT03283384

Clonal evolution analysis along with RNA-seq data point to the role of genomic instability and lineage switching in driving acquired resistance. Specifically, the activity of the HH pathway may play a critical role in resistance to ARPI.

The analyses revealed important takeaways: 1) key activating mutations can be identified using a simple liquid biopsy, 2) some embryonal RMS cases have PD-L1 expression, and 3) OncoSignal analysis may be useful in identifying oncogenic driver pathways. These findings helped to identify that this child was eligible for a clinical trial that contained combined MEK and PD-L1 checkpoint inhibitors, a potential solution to treating embryonal RMS.

Conclusions : Molecular subtyping identified tumors from high-risk NMIBC patients with an aggressive phenotype and a poor response to BCG. Our findings provide a clinical tool for improved identification of high-risk NMIBC patients at high risk of progression, which can be used to select patients for early radical cystectomy or to develop novel subtype-directed therapies.

"Protean BioDiagnostics presents a new method to identify potential therapeutic targets in breast cancer...The method called OncoSignal analysis utilizes mRNA transcriptional measurements to accurately calculate the pathway activity of seven key oncogenic signaling pathways."

OncoSignal™ analysis identifies enhanced targetable oncogenic pathway activity in a majority of TNBC breast cancers. Of interest, 7 of 88 cases (8%) classified as TNBC using IHC methods showed evidence of estrogen receptor signal pathway activation, and 15 (17%) showed elevated AR PAS. PI3K and MAPK had high PAS in over 85% of TNBC cases.