GAB1-ABL1 fusions in tumors that have histologic overlap with NTRK-rearranged spindle cell tumors.

Fibroblastic spindle cell tumors are a heterogeneous group of rare soft tissue tumors that are increasingly recognized as associated with a variety of kinase gene fusions. We report two cases of GAB1-ABL1 fusions in spindle cell tumors that histologically overlap with neurotrophic tyrosine receptor kinase (NTRK)-rearranged spindle cell tumors. The first case occurred in a 76-year-old female who had a large deep-seated spindle cell tumor composed of monotonous ovoid to spindle cells in a background of thick stromal collagen bands with prominent hyalinized vessels and inconspicuous mitoses (<1/10 HPF). Immunohistochemical stains showed co-expression of S100 and CD34. A GAB1-ABL1 fusion was detected by whole transcriptome RNA sequencing. The patient had a partial response to imatinib. The second case was previously described as a solitary fibrous tumor, occurring in a 9-year-old female with a cellular spindle cell tumor with patchy CD34 immunoexpression but no expression of S100. Upon clinicopathologic re-review, including anchored multiplex next-generation sequencing, a GAB1-ABL1 fusion was identified. In summary, we report the first two cases of spindle cell tumors with variable expression of CD34 and/or S100, driven by GAB1-ABL1 gene fusions with histologic overlap with NTRK-rearranged spindle cell tumors, suggesting that ABL-fusions may also be oncogenic drivers within this spectrum of tumors. These cases highlight the evolving understanding of fibroblastic spindle cell tumor biology and the utility of sequencing in identifying a targetable alteration.

Functional impact and targetability of PI3KCA, GNAS, and PTEN mutations in a spindle cell rhabdomyosarcoma with MYOD1 L122R mutation.

Spindle cell/sclerosing rhabdomyosarcoma (ssRMS) is a rare subtype of rhabdomyosarcoma, commonly harboring a gain-of-function L122R mutation in the muscle-specific master transcription factor MYOD1. MYOD1-mutated ssRMS is almost invariably fatal, and development of novel therapeutic approaches based on the biology of the disease is urgently needed. MYOD1 L122R affects the DNA-binding domain and is believed to confer MYC-like properties to MYOD1, driving oncogenesis. Moreover, the majority of the MYOD1-mutated ssRMS harbor additional alterations activating the PI3K/AKT pathway. It is postulated that the PI3K/AKT pathway cooperates with MYOD1 L122R. To address this biological entity, we established and characterized a new patient-derived ssRMS cell line OHSU-SARC001, harboring MYOD1 L122R as well as alterations in PTEN, PIK3CA, and GNAS We explored the functional impact of these aberrations on oncogenic signaling with gain-of-function experiments in C2C12 murine muscle lineage cells. These data reveal that PIK3CAI459_T462del, the novel PIK3CA variant discovered in this patient specimen, is a constitutively active kinase, albeit to a lesser extent than PI3KCAE545K, a hotspot oncogenic mutation. Furthermore, we examined the effectiveness of molecularly targeted PI3K/AKT/mTOR and RAS/MAPK inhibitors to block oncogenic signaling and suppress the growth of OHSU-SARC001 cells. Dual PI3K/mTOR (LY3023414, bimiralisib) and AKT inhibitors (ipatasertib, afuresertib) induced dose-dependent reductions in cell growth. However, mTOR-selective inhibitors (everolimus, rapamycin) alone did not exert cytotoxic effects. The MEK1/2 inhibitor trametinib did not impact proliferation even at the highest doses tested. Our data suggest that molecularly targeted strategies may be effective in PI3K/AKT/mTOR-activated ssRMS. Taken together, these data highlight the importance of utilizing patient-derived models to assess molecularly targetable treatments and their potential as future treatment options.

Diagnosing water treatment critical control points for cyanobacterial removal: Exploring benefits of combined microscopy, next-generation sequencing, and cell integrity methods.

A wide range of cyanobacterial species and their harmful metabolites are increasingly detected in water bodies worldwide, exacerbated by climate change and human activities. The resulting bloom conditions represent significant challenges to production of safe drinking water and cost effective water reuse, therefore their removal is a priority to ensure public safety. While current microscopic taxonomy identification methods provide valuable information about cell numbers during treatment, these methods are incapable of providing information about the fate of cells during treatment. The objectives of this study were to (1) identify the critical control points for breakthrough and accumulation of cells by investigating the fate of cells during treatment processes using a combination of taxonomy, cell integrity and next-generation sequencing (NGS), and (2) assess the impact of pre-treatment processes on breakthrough prevention at critical control points, and the benefits of cell integrity and NGS analysis for improved management purposes. This paper presents the results of an unprecedented cyanobacterial monitoring program conducted in four full scale water treatment plants located in three different climate zones. Cyanobacterial cell integrity and accumulation during operation process were assessed for the first time using next generation of gene sequencing methods. NGS analysis led to detection of cyanobacterial and melainabacteria orders in water samples that were not identified by microscopy. 80 ±â€¯5% of cells were completely lysed post pre-oxidation (for both ozone and potassium permanganate). However unlike pre-ozonation, the remaining cells were undamaged cells with the potential to accumulate and grow within the plants post-KMnO4 treatment, particularly in clarifier sludge. To effectively monitor water quality, this study presents a synergistic approach coupling new and traditional analytical methods and demonstrates the importance of identifying critical points for managing accumulation of cyanobacteria within plants.