TEST:

VENTANA ALK (D5F3) CDx Assay

P2, N=109, Completed, Pfizer | Active, not recruiting --> Completed

Screening for ALK rearrangements with IHC is both reliable and cost effective. In-house testing with IHC removes reliance on send-out testing for ALK detection, effectively cutting costs and decreasing TAT. Additionally, the creation of a reflex algorithm can improve lung cancer biomarker testing processes within hospitals.

These findings further substantiate the hypothesis that SAMS and epithelioid FH may represent a morphologic and genetic spectrum. The differential diagnosis, clinical follow up, and the molecular genetic findings will be discussed.

P3; Active, not recruiting --> Completed

FISH testing can be technically challenging and difficult to interpret. NSCLC with atypical/abnormal FISH pattern is a rare event that should be considered positive for ALK rearrangement and should benefit from ALK-targeted therapy. This study highlights the continued discontinuation of the use and compatibility of immunohistochemistry as a preliminary screening method or confirmation of atypical cases by alternative techniques such as next-generation sequencing.

This study demonstrates that strong but focal ALK (D5F3) immunostaining and strong expression in neuroendocrine differentiation may not indicate ALK fusion. By considering these findings, we can improve the accuracy of patient selection for targeted therapy by minimizing false-positive interpretations of ALK (D5F3) staining.

P3; Trial primary completion date: Aug 2024 --> Feb 2024

Positive protein expression regardless of FISH result correlated more with a positive NGS result compared to samples with a positive FISH result with negative protein expression. FISH analysis was able to detect atypical or heterogenous patterns of rearrangement in a proportion of cases with negative protein expression, which may be associated with more extensive genetic alterations rather than true ALK rearrangement.

Blood glucose control also has a far-reaching impact on the occurrence, development and prognosis of lung cancer. Attention should be paid to the follow-up of the above-mentioned patients.

Among them, immunohistochemistry (IHC)(Ventana D5F3), RNA based next-generation sequencing (RNA-based NGS) confirmed positive echinoderm microtubule associated protein like 4 (EML4)-ALK fusion, while DNA-based NGS was negative. This paper analyzed the detection methods of ALK fusion, in order to clarify which detection method is the most accurate and simple to choose in different clinical cases and guide the subsequent treatment..

This study highlights the real-world variability and challenges of ALK test in advanced NSCLC, demonstrating a predominant use of IHC-VENTANA-D5F3 with high consistency and distinct clinicopathological features in ALK-positive patients. These findings underscore the need for a consensus on optimal test practices and support the development of refined ALK test strategies to enhance diagnostic accuracy and therapeutic decision-making in NSCLC.

Surgical resection has been successful in treating cutaneous disease, with no reported recurrence. As further cases of this rare condition come to light, its full clinicopathological and molecular features may become more apparent.

P3; Trial completion date: May 2024 --> Sep 2024 | Trial primary completion date: Apr 2024 --> Aug 2024

P2 | N=33 | Active, not recruiting | Sponsor: Fundación GECP | Trial primary completion date: Apr 2024 ➔ Dec 2024

This is the largest cohort from India reporting real world uptake of NGS based biomarker testing and targeted therapy. There is a clear disparity in terms of gender and age group, possibly attributable to social stigmata and taboos prevailing in rural parts of the country. Despite increased detection rates, access to targeted therapy is still a concern owing to inaccessibility to drugs/ lack of global clinical trials in India as well as lack of patient support programs/groups.

Our inhouse panel was associated with rapid TAT and faster initiation of systemic treatment. Approximately 30% of patients would not need tissue sent for NGS based on inhouse detection of a potentially actionable mutation.

The NGS testing revealed EML4::ALK fusion in three, STRN::ALK fusion in two, and ITSN2::ALK fusion in one. In conclusion, although ALK-rearranged PTCs have been associated with neutral gene expression profile from a BRAF-RAS scoring perspective, the "RAS-like" nuclear features were more commonly identified in this series, resulting in frequent indeterminate diagnosis of preoperative FNA.

ALK was highly expressed in NB. ALK expression was not related to several clinical and histopathological features. More studies are needed to elucidate the association between ALK expression and ALK gene status and to investigate disease progression, especially the oncogenesis of ALK-positive NB.

P1/2; N=12; Recruiting; Sponsor:University of Colorado, Denver

Immunostaining is a robust method for ALK-rearrangement testing, with fluorescence in situ hybridization adding value in the rare equivocal stained case. ROS1-rearrangement testing is more cost-effective if immunohistochemistry is followed by fluorescence in situ hybridization after excluding EGFR-mutant and ALK-rearranged adenocarcinomas.

Following corticosteroid treatment and discontinuation of alectinib, clinical presentations and CT scan gradually improved, but the primary lung lesions enlarged in the regular follow-up. The administration of a first-generation ALK inhibitor crizotinib was then initiated and the disease was stable for 25 months without recurrence of ILD.

Immunohistochemistry is a useful tool for confirming the Spitz histogenesis of a melanocytic tumour when it demonstrates the presence of expression of a kinase fusion. Our data suggest that the ALK-1 antibody is significantly less sensitive than ALK-D5F3 and ALK-5A4 antibodies for the detection of ALK-rearranged Spitz tumors and we do not recommend its use for this purpose.

Immunohistochemistry-based assays provide a valid and reasonably priced alternative, especially in settings where molecular confirmatory tests are neither offered nor accessible. Given high interassay and molecular concordance, we propose that the novel Dako OTI1A4 assay can be reliably used to identify cases with ALK rearrangement.

Conclusion Our study confirms that compared to IHC, NGS is a more accurate first line testing platform for the detection of consensus ALK fusion status. However access to IHC may be required to mitigate the higher tissue requirements of NGS and access to FISH may be beneficial to resolve atypical results.

P1/2; Not yet recruiting --> Recruiting | Initiation date: Dec 2023 --> Jul 2023

Gene co-occurrence survival analysis showed that mPFS of pts harboring FAT3/FAT4 mutations were significantly shorter than those without FAT3/FAT4 mutations (8.9 mo vs 16.5 mo, p=0.012). Conclusions Clinical utility of ctDNA was demonstrated, not only at baseline by identifying fusion types and subgroups of ALK+ NSCLC pts that may benefit more from SAF-189s, but also at progression by tracking ALK-resistant mutations.

Ventana ALK (D5F3) immunohistochemistry improved the accuracy of pathologic assessment after neoadjuvant ALK-TKIs for ALK-positive NSCLC. It is recommended to use Ventana ALK D5F3 immunohistochemistry to help confirm MPR and PCR in future pathological assessment work.

In this regard, Dako-CD246 proves to yield concordant results with Ventana-D5F3, which is already promptly used in routine pathology practice. The present study being the first in the field, given its high inter-assay and molecular concordance, we propose that novel Dako-CD246 assay can be reliably used in identifying cases for ALK-targeted therapy.

P3; Trial primary completion date: Apr 2023 --> Apr 2024

P1/2; N=12; Not yet recruiting; Sponsor:University of Colorado, Denver

P2; Trial completion date: Aug 2023 --> Dec 2024

In this case series, all six ALK -rearranged PTCs demonstrated a predominant follicular growth pattern and five of them were diagnosed as infiltrative follicular variant. The nuclear features were subtle and might be present in only focal areas, resulting in a significant rate of false negative diagnosis (33%, 2/6) in pre-operative FNA.

We identified a subset of microcystic/reticular schwannomas with NONO::TFE3 fusions. TFE3 and ALK-D5F3 immunohistochemistry appear to be reliable surrogates for fusion status.

P3; Recruiting --> Active, not recruiting

P2; Trial completion date: Aug 2022 --> Aug 2023

In Taiwan, the patients with thoracic IMT were young, and had early stages of disease status. ALK rearrangement and NTRK3 overexpression were the major genetic alterations. Different partners of ALK arrangement could be detected.

ALK-positive patients experience clinical benefit from pemetrexed-based chemotherapy possibly due to lower thymidylate synthase (TS) levels...D5F3 and 5A4 clones have the highest percentage of agreement. TS levels are significantly lower in FISH-positive patients.

An Archer FusionPlex panel assay demonstrated a fusion involving both exons 4 from the JAZF1 and BCORL1 genes. The JAZF1::BCORL1 has not, to the best of our knowledge, been previously reported in a benign/low-grade mesenchymal uterine lesion.

P2; Active, not recruiting --> Completed | Trial completion date: May 2022 --> Jul 2021

Patients with advanced ALK-positive (by immunohistochemistry) NSCLC were randomized 1:1 to alectinib 600 mg or crizotinib 250 mg, twice daily. Reasonable concordance between FACT and immunohistochemistry was observed; both methods are valuable in identifying ALK-positive patients, separately or concurrently. Alectinib was found to have superior PFS in the plasma ALK-positive population, as in the ITT population.

P2; Recruiting --> Active, not recruiting

"Moreover, the patient achieved an admirable response to alectinib, with a clinical evaluation of complete response (CR). In summary, our findings expand the spectrum of ALK fusion patterns and provide robust evidence for the precise administration of alectinib in the future."