Conventional and semi-automatic histopathological analysis of tumor cell content for multigene sequencing of lung adenocarcinoma

Kazdal, Daniel; Rempel, Eugen; Oliveira, Cristiano; Allgäuer, Michael; Harms, Alexander; Singer, Kerstin; Kohlwes, Elke; Ormanns, Steffen; Fink, Ludger; Kriegsmann, Jörg; Leichsenring, Michael; Kriegsmann, Katharina; Stögbauer, Fabian; Tavernar, Luca; Leichsenring, Jonas; Volckmar, Anna‐Lena; Longuespée, Rémi; Winter, Hauke; Eichhorn, Martin; Herth, Felix J.F.; Christopoulos, Petros; Reck, Martin; Muley, Thomas; Weichert, Wilko; Budczies, Jan; Thomas, Michael; Peters, Solange; Warth, Arne; Schirmacher, Peter; Kriegsmann, Mark

doi:10.21037/tlcr-20-1168

Cited by 8 publications

(4 citation statements)

References 25 publications

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“…Yamaguchi et al [9] described that residual LBC samples can be directly used for DNA extraction and NGS analysis provided they contain sufficient amounts of tumor fraction. Therefore, to increase the probability of the success, tumor cellularity or tumor cell content should be considered as a crucial process of the preanalytical stage [27]. LBC is advantageous in providing better quality materials with less contamination than conventional cytologic preparations and serves as a source for molecular testing [1,28].…”

Section: Discussionmentioning

confidence: 99%

Optimal Preservations of Cytological Materials Using Liquid-Based Cytology Fixatives for Next-Generation Sequencing Analysis

et al. 2022

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Introduction: Molecular targeted therapies have been established for various diseases, including cancers, and there is an increasing need for molecular testing on cytology specimens. The aim of this study was to determine the optimal preservation methods of liquid-based cytology (LBC) materials for molecular testing. Methods: Cytological samples from 35 surgical resected non-small cell lung carcinoma specimens were obtained between June 2016 and June 2021. The samples were fixed in CytoRich™ red Preservative and stored at 4°C. One week later, three tubes were prepared from each specimen sample and divided into the following groups: the SurePath™ group (continued storage at 4°C), Frozen (Fr) group (stored at −80°C after centrifugation), and LBC-Cell Block (LBC-CB) group (generation of paraffin-embedded CB and storage at 4°C). Samples from 5 patients were used for the time course analysis, and we performed evaluations on these samples at 1, 3, 6, 12, 24, and 36 months. The concentrations and purities of extracted DNA and RNA were measured. The double-stranded DNA (dsDNA) and RNA concentrations were also measured by a fluorometer. The DNA and RNA integrities were quantified by the DNA and RNA integrity number. Results: Evaluation of samples was performed at baseline and the six timepoints. In the LBC-CB group, DNA and dsDNA concentrations were higher rather than those in the other groups. The RNA concentration of the LBC-CB group was relatively high compared with those of the other groups at the 36-month timepoint. The Fr group maintained higher DNA quality compared with the other groups over 3 years. The LBC-CB group maintained a higher RNA quality than the other groups until 24 months. Conclusion: LBC-CB preparation is an effective method to maintain DNA/RNA quality and quantity in long-duration preservation for eventual molecular testing. Therefore, LBC-CB may have applications on preanalytical stage for molecular genomic testing such as next-generation sequencing.

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Section: Discussionmentioning

confidence: 99%

Optimal Preservations of Cytological Materials Using Liquid-Based Cytology Fixatives for Next-Generation Sequencing Analysis

et al. 2022

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“…Digital image analysis was performed by an experienced pathologist in a semiautomatic manner using QuPath, a more detailed description can be found in the online methods [ 20 , 21 ].…”

Section: Methodsmentioning

confidence: 99%

Accurate tumor purity determination is critical for the analysis of homologous recombination deficiency (HRD)

Menzel

Endris

Schwab

et al. 2023

Translational Oncology

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“…As mentioned above, digital pathology will certainly and progressively enhance or replace the present practices [ 77 , 78 , 92 – 95 ]. In addition to improving the diagnostic processes, digital pathology can compile many information and imaging data.…”

Section: Short- and Mid-term Perspectivesmentioning

confidence: 99%

Current challenges and practical aspects of molecular pathology for non-small cell lung cancers

Hofman,

Berezowska,

Kazdal

et al. 2023

Virchows Arch

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The continuing evolution of treatment options in thoracic oncology requires the pathologist to regularly update diagnostic algorithms for management of tumor samples. It is essential to decide on the best way to use tissue biopsies, cytological samples, as well as liquid biopsies to identify the different mandatory predictive biomarkers of lung cancers in a short turnaround time. However, biological resources and laboratory member workforce are limited and may be not sufficient for the increased complexity of molecular pathological analyses and for complementary translational research development. In this context, the surgical pathologist is the only one who makes the decisions whether or not to send specimens to immunohistochemical and molecular pathology platforms. Moreover, the pathologist can rapidly contact the oncologist to obtain a new tissue biopsy and/or a liquid biopsy if he/she considers that the biological material is not sufficient in quantity or quality for assessment of predictive biomarkers. Inadequate control of algorithms and sampling workflow may lead to false negative, inconclusive, and incomplete findings, resulting in inappropriate choice of therapeutic strategy and potentially poor outcome for patients. International guidelines for lung cancer treatment are based on the results of the expression of different proteins and on genomic alterations. These guidelines have been established taking into consideration the best practices to be set up in clinical and molecular pathology laboratories. This review addresses the current predictive biomarkers and algorithms for use in thoracic oncology molecular pathology as well as the central role of the pathologist, notably in the molecular tumor board and her/his participation in the treatment decision-making. The perspectives in this setting will be discussed.

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Conventional and semi-automatic histopathological analysis of tumor cell content for multigene sequencing of lung adenocarcinoma

Cited by 8 publications

References 25 publications

Optimal Preservations of Cytological Materials Using Liquid-Based Cytology Fixatives for Next-Generation Sequencing Analysis

Optimal Preservations of Cytological Materials Using Liquid-Based Cytology Fixatives for Next-Generation Sequencing Analysis

Accurate tumor purity determination is critical for the analysis of homologous recombination deficiency (HRD)

Current challenges and practical aspects of molecular pathology for non-small cell lung cancers

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