HIGH THROUGHPUT TCR SEQUENCING PROVIDES ADDED VALUE IN THE DIAGNOSIS OF CUTANEOUS T-CELL LYMPHOMA
(Abstract release date: 05/19/16)
EHA Library. Kirsch I. 06/09/16; 132697; E1148
Dr. Ilan Kirsch
Contributions
Contributions
Abstract
Abstract: E1148
Type: Eposter Presentation
Background
Diagnosis of early-stage CTCL can be challenging because skin lesions contain a mixture of both diverse benign and clonal malignant T cells. High throughput sequencing (HTS) of the TCR gamma (TCRG) and TCR beta (TCRB) CDR3 regions is a technique that provides comprehensive analysis of the total repertoire of T cell clones in a specimen, the breadth of repertoire diversity, and an exact quantitation of individual T cell clone
Aims
We investigated whether the Adaptive Biotechnologies multiplex PCR and high throughput sequencing assays of TCRB and TCRG could discriminate between benign and malignant T-cell skin dyscrasias
Methods
We analyzed skin biopsies from 46 patients with histopathologically and clinically (based on presentation, history, and course) labelled CTCL (over 80% Stage I or Stage II), 23 patients with psoriasis, 11 eczematous dermatitis, 10 allergic dermatitis, and 6 with normal skin findings.
Results
The TCRB and TCRG assays were able to define the dominant clonal sequences in every CTCL case and to distinguish likely gamma/delta from alpha/beta lymphoma. Using the fraction of the top two TCRG sequences as a fraction of the total nucleated cell population defined a cut off of approximately 1/1000 above which the biopsy was highly specific for malignant disease and below which the assay approached 100% specificity for non-malignant disease. The discrimination afforded by reference to the per cent comprised by the top two TCRG sequences of all TCRG sequences generated from the sample (most comparable to data generated by TCR PCR) was not nearly as robust. TCR PCR failed to provide relevant categorization in approximately 30% of the same CTCL cases. This approach also was able to identify low level presence of CTCL-correlated clones in systemic circulation and to distinguish subsequent inflammatory reactions from disease recurrence. Analysis of CTCL TCRG genes was consistent with CTCL being a malignancy derived from mature T cells.
Conclusion
Multiplex PCR and high throughput sequencing of TCRG and TCRB loci in suspect skin lesions was able to impact and add value to the diagnosis and monitoring of patients with early stage cutaneous T-cell lymphoma.
Session topic: E-poster
Keyword(s): Cutaneous T-cell lymphoma, Diagnosis, TCR
Type: Eposter Presentation
Background
Diagnosis of early-stage CTCL can be challenging because skin lesions contain a mixture of both diverse benign and clonal malignant T cells. High throughput sequencing (HTS) of the TCR gamma (TCRG) and TCR beta (TCRB) CDR3 regions is a technique that provides comprehensive analysis of the total repertoire of T cell clones in a specimen, the breadth of repertoire diversity, and an exact quantitation of individual T cell clone
Aims
We investigated whether the Adaptive Biotechnologies multiplex PCR and high throughput sequencing assays of TCRB and TCRG could discriminate between benign and malignant T-cell skin dyscrasias
Methods
We analyzed skin biopsies from 46 patients with histopathologically and clinically (based on presentation, history, and course) labelled CTCL (over 80% Stage I or Stage II), 23 patients with psoriasis, 11 eczematous dermatitis, 10 allergic dermatitis, and 6 with normal skin findings.
Results
The TCRB and TCRG assays were able to define the dominant clonal sequences in every CTCL case and to distinguish likely gamma/delta from alpha/beta lymphoma. Using the fraction of the top two TCRG sequences as a fraction of the total nucleated cell population defined a cut off of approximately 1/1000 above which the biopsy was highly specific for malignant disease and below which the assay approached 100% specificity for non-malignant disease. The discrimination afforded by reference to the per cent comprised by the top two TCRG sequences of all TCRG sequences generated from the sample (most comparable to data generated by TCR PCR) was not nearly as robust. TCR PCR failed to provide relevant categorization in approximately 30% of the same CTCL cases. This approach also was able to identify low level presence of CTCL-correlated clones in systemic circulation and to distinguish subsequent inflammatory reactions from disease recurrence. Analysis of CTCL TCRG genes was consistent with CTCL being a malignancy derived from mature T cells.
Conclusion
Multiplex PCR and high throughput sequencing of TCRG and TCRB loci in suspect skin lesions was able to impact and add value to the diagnosis and monitoring of patients with early stage cutaneous T-cell lymphoma.
Session topic: E-poster
Keyword(s): Cutaneous T-cell lymphoma, Diagnosis, TCR
Abstract: E1148
Type: Eposter Presentation
Background
Diagnosis of early-stage CTCL can be challenging because skin lesions contain a mixture of both diverse benign and clonal malignant T cells. High throughput sequencing (HTS) of the TCR gamma (TCRG) and TCR beta (TCRB) CDR3 regions is a technique that provides comprehensive analysis of the total repertoire of T cell clones in a specimen, the breadth of repertoire diversity, and an exact quantitation of individual T cell clone
Aims
We investigated whether the Adaptive Biotechnologies multiplex PCR and high throughput sequencing assays of TCRB and TCRG could discriminate between benign and malignant T-cell skin dyscrasias
Methods
We analyzed skin biopsies from 46 patients with histopathologically and clinically (based on presentation, history, and course) labelled CTCL (over 80% Stage I or Stage II), 23 patients with psoriasis, 11 eczematous dermatitis, 10 allergic dermatitis, and 6 with normal skin findings.
Results
The TCRB and TCRG assays were able to define the dominant clonal sequences in every CTCL case and to distinguish likely gamma/delta from alpha/beta lymphoma. Using the fraction of the top two TCRG sequences as a fraction of the total nucleated cell population defined a cut off of approximately 1/1000 above which the biopsy was highly specific for malignant disease and below which the assay approached 100% specificity for non-malignant disease. The discrimination afforded by reference to the per cent comprised by the top two TCRG sequences of all TCRG sequences generated from the sample (most comparable to data generated by TCR PCR) was not nearly as robust. TCR PCR failed to provide relevant categorization in approximately 30% of the same CTCL cases. This approach also was able to identify low level presence of CTCL-correlated clones in systemic circulation and to distinguish subsequent inflammatory reactions from disease recurrence. Analysis of CTCL TCRG genes was consistent with CTCL being a malignancy derived from mature T cells.
Conclusion
Multiplex PCR and high throughput sequencing of TCRG and TCRB loci in suspect skin lesions was able to impact and add value to the diagnosis and monitoring of patients with early stage cutaneous T-cell lymphoma.
Session topic: E-poster
Keyword(s): Cutaneous T-cell lymphoma, Diagnosis, TCR
Type: Eposter Presentation
Background
Diagnosis of early-stage CTCL can be challenging because skin lesions contain a mixture of both diverse benign and clonal malignant T cells. High throughput sequencing (HTS) of the TCR gamma (TCRG) and TCR beta (TCRB) CDR3 regions is a technique that provides comprehensive analysis of the total repertoire of T cell clones in a specimen, the breadth of repertoire diversity, and an exact quantitation of individual T cell clone
Aims
We investigated whether the Adaptive Biotechnologies multiplex PCR and high throughput sequencing assays of TCRB and TCRG could discriminate between benign and malignant T-cell skin dyscrasias
Methods
We analyzed skin biopsies from 46 patients with histopathologically and clinically (based on presentation, history, and course) labelled CTCL (over 80% Stage I or Stage II), 23 patients with psoriasis, 11 eczematous dermatitis, 10 allergic dermatitis, and 6 with normal skin findings.
Results
The TCRB and TCRG assays were able to define the dominant clonal sequences in every CTCL case and to distinguish likely gamma/delta from alpha/beta lymphoma. Using the fraction of the top two TCRG sequences as a fraction of the total nucleated cell population defined a cut off of approximately 1/1000 above which the biopsy was highly specific for malignant disease and below which the assay approached 100% specificity for non-malignant disease. The discrimination afforded by reference to the per cent comprised by the top two TCRG sequences of all TCRG sequences generated from the sample (most comparable to data generated by TCR PCR) was not nearly as robust. TCR PCR failed to provide relevant categorization in approximately 30% of the same CTCL cases. This approach also was able to identify low level presence of CTCL-correlated clones in systemic circulation and to distinguish subsequent inflammatory reactions from disease recurrence. Analysis of CTCL TCRG genes was consistent with CTCL being a malignancy derived from mature T cells.
Conclusion
Multiplex PCR and high throughput sequencing of TCRG and TCRB loci in suspect skin lesions was able to impact and add value to the diagnosis and monitoring of patients with early stage cutaneous T-cell lymphoma.
Session topic: E-poster
Keyword(s): Cutaneous T-cell lymphoma, Diagnosis, TCR
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