Utility of flow cytometry and gene rearrangement analysis in tissue and blood of patients with suspected cutaneous T‑cell lymphoma

Gibbs,Julie D.; Ma,Sophia; Kim,Anna; Seminario‑Vidal,Lucia; Sokol,Lubomir; Zhang,Hailing; Zhang,Xiaohui; Sagatys,Elizabeth; Chen,Pei‑Ling; Messina,Jane L.

doi:10.3892/or.2020.7865

Utility of flow cytometry and gene rearrangement analysis in tissue and blood of patients with suspected cutaneous T‑cell lymphoma

Authors:
- Julie D. Gibbs
- Sophia Ma
- Anna Kim
- Lucia Seminario‑Vidal
- Lubomir Sokol
- Hailing Zhang
- Xiaohui Zhang
- Elizabeth Sagatys
- Pei‑Ling Chen
- Jane L. Messina
View Affiliations

Affiliations: Bay Pines VA Medical Center, St. Petersburg, FL 33744, USA, Sutter Health, San Ramon, CA 94583, USA, Department of Dermatology and Cutaneous Surgery, University of South Florida Morsani College of Medicine, Tampa, FL 33612, USA, Department of Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA, Department of Hematopathology and Laboratory Medicine, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA, Department of Anatomic Pathology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA
Published online on: November 23, 2020 https://doi.org/10.3892/or.2020.7865
Pages: 349-358

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cutaneous T‑cell lymphoma (CTCL) is difficult to diagnose at an early stage. Current diagnostic tools include clinical examination, histomorphologic analysis, immunohistochemistry, flow cytometry of peripheral blood and/or lesional tissue, and evaluation of T‑cell receptor (TCR) clonality by gene rearrangement analysis (TCRGR). Advances in genomic sequencing, including high‑throughput sequencing (HTS), can be used to identify specific clones of rearranged TCR genes. Even with all of these tools, CTCL can take as long as a decade to definitively diagnose, potentially delaying treatment options and causing patient anxiety. This study aimed to evaluate the performance of the various ancillary testing modalities used to diagnose early‑stage CTCL. In a subset of patients the performance of HTS was compared to flow cytometry and conventional TCRGR analysis via polymerase chain reaction (PCR). Fifty‑five patients, with a total of 68 skin biopsies and peripheral blood draws, were evaluated using flow cytometry, PCR‑TCRGR, and HTS‑TCRGR to determine the sensitivity and specificity of each ancillary test. In tissue biopsies, flow cytometry (64%), PCR (71%) and HTS (69%) shared similar sensitivities; flow cytometry had the highest specificity (93%), followed by HTS (86%) and PCR (76.9%). However, flow cytometry and PCR had insufficient DNA quantities in 29 and 15% of the specimens, respectively. Peripheral blood testing was less sensitive than tissue testing (flow cytometry 14%, PCR 41%, HTS 33%); in peripheral blood, HTS was the most specific test (flow cytometry, 70%; PCR, 62.5%; and HTS, 100%). HTS is a highly specific molecular test for identifying CTCL in peripheral blood and skin biopsy specimens; however, our findings suggest a need for a continued search for more sensitive tests for early‑stage CTCL.

View Figures

View References

1	Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H and Thiele J: World Health Organization Classification of Tumours of Hematopoietic and Lymphoid Tissue. IARC Press; Lyon: 2008
2	Cerroni L: Lymphoproliferative lesions of the skin. J Clin Pathol. 59:813–826. 2006. View Article : Google Scholar
3	Willemze R and Meijer CJ: Classification of cutaneous T-cell lymphoma: From Alibert to WHO-EORTC. J Cutan Pathol. 33 (Suppl 1):18–26. 2006. View Article : Google Scholar
4	Olsen EA: Evaluation, diagnosis, and staging of cutaneous lymphoma. Dermatol Clin. 33:643–654. 2015. View Article : Google Scholar
5	Algara P, Soria C, Martinez P, Sanchez L, Villuendas R, Garcia P, Lopez C, Orradre JL and Piris MA: Value of PCR detection of TCR gamma gene rearrangement in the diagnosis of cutaneous lymphocytic infiltrates. Diagn Mol Pathol. 3:275–282. 1994. View Article : Google Scholar
6	Galper SL, Smith BD and Wilson LD: Wilson, Diagnosis and management of mycosis fungoides. Oncology (Williston Park). 24:491–501. 2010.
7	Willemze R, Jaffe ES, Burg G, Cerroni L, Berti E, Swerdlow SH, Ralfkiaer E, Chimenti S, Diaz-Perez JL, Duncan LM, et al: WHO-EORTC classification for cutaneous lymphomas. Blood. 105:3768–3785. 2005. View Article : Google Scholar
8	Cerroni L: Mycosis fungoides-clinical and histopathologic features, differential diagnosis, and treatment. Semin Cutan Med Surg. 37:2–10. 2018. View Article : Google Scholar
9	Jawed SI, Myskowski PL, Horwitz S, Moskowitz A and Querfeld C: Primary cutaneous T-cell lymphoma (mycosis fungoides and Sézary syndrome): Part I. Diagnosis: Clinical and histopathologic features and new molecular and biologic markers. J Am Acad Dermatol. 70:205.e1–16; quiz 221-222. 2014. View Article : Google Scholar
10	Kempf W and Mitteldorf C: Mitteldorf, pathologic diagnosis of cutaneous lymphomas. Dermatol Clin. 33:655–681. 2015. View Article : Google Scholar
11	Hodak E and Amitay-Laish I: Amitay-Laish, Mycosis fungoides: A great imitator. Clin Dermatol. 37:255–267. 2019. View Article : Google Scholar
12	Campbell SM, Peters SB, Zirwas MJ and Wong HK: Immunophenotypic diagnosis of primary cutaneous lymphomas: A review for the practicing dermatologist. J Clin Aesthet Dermatol. 3:21–25. 2010.
13	Ashton-Key M, Diss TC, Du MQ, Kirkham N, Wotherspoon A and Isaacson PG: The value of the polymerase chain reaction in the diagnosis of cutaneous T-cell infiltrates. Am J Surg Pathol. 21:743–747. 1997. View Article : Google Scholar
14	Alaibac M, Pigozzi B, Belloni-Fortina A, Michelotto A, Saponeri A and Peserico A: CD7 expression in reactive and malignant human skin T-lymphocytes. Anticancer Res. 23:2707–2710. 2003.
15	Moll M, Reinhold U, Kukel S, Abken H, Müller R, Oltermann I and Kreysel HW: CD7-negative helper T cells accumulate in inflammatory skin lesions. J Invest Dermatol. 102:328–332. 1994. View Article : Google Scholar
16	Lu C, Zhang J, Nagahawatte P, Easton J, Lee S, Liu Z, Ding L, Wyczalkowski MA, Valentine M, Navid F, et al: The genomic landscape of childhood and adolescent melanoma. J Invest Dermatol. 135:816–823. 2015. View Article : Google Scholar
17	Skov AG and Gniadecki R: Gniadecki, Delay in the histopathologic diagnosis of mycosis fungoides. Acta Derm Venereol. 95:472–475. 2015. View Article : Google Scholar
18	van Doorn R, Van Haselen CW, van Voorst Vader PC, Geerts ML, Heule F, de Rie M, Steijlen PM, Dekker SK, van Vloten WA and Willemze R: Mycosis fungoides: Disease evolution and prognosis of 309 Dutch patients. Arch Dermatol. 136:504–510. 2000. View Article : Google Scholar
19	Kirsch IR, Watanabe R, O'Malley JT, Williamson DW, Scott LL, Elco CP, Teague JE, Gehad A, Lowry EL, LeBoeuf NR, et al: TCR sequencing facilitates diagnosis and identifies mature T cells as the cell of origin in CTCL. Sci Transl Med. 7:308ra1582015. View Article : Google Scholar
20	Maitre E, Le-Page AL, Comoz F, Truquet F, Damaj G, Cornet E, Verneuil L, Salaün V and Troussard X: Usefulness of flow cytometry for the detection of cutaneous localization in malignant hematologic disorders. Cytometry B Clin Cytom. 96:283–293. 2019. View Article : Google Scholar
21	Bawazir MA, Almohideb M, Walsh S, Shear NH and Alhusayen R: Early-stage mycosis fungoides screening investigations: A retrospective analysis of 440 cases. J Eur Acad Dermatol Venereol. 32:e217–e218. 2018. View Article : Google Scholar
22	Horna P, Kurant D, Sokol L, Sotomayor EM, Moscinski L and Glass LF: Flow cytometric identification of immunophenotypically aberrant T-cell clusters on skin shave biopsy specimens from patients with mycosis fungoides. Am J Clin Pathol. 143:785–796. 2015. View Article : Google Scholar
23	Rea B, Haun P, Emerson R, Vignali M, Farooqi M, Samimi S, Elenitsas R, Kirsch I and Bagg A: Role of high-throughput sequencing in the diagnosis of cutaneous T-cell lymphoma. J Clin Pathol. 71:814–820. 2018. View Article : Google Scholar
24	de Masson A, O'Malley JT, Elco CP, Garcia SS, Divito SJ, Lowry EL, Tawa M, Fisher DC, Devlin PM and Teague JE: High-throughput sequencing of the T cell receptor β gene identifies aggressive early-stage mycosis fungoides. Sci Transl Med. 10:eaar58942018. View Article : Google Scholar
25	Weng WK, Armstrong R, Arai S, Desmarais C, Hoppe R and Kim YH: Minimal residual disease monitoring with high-throughput sequencing of T cell receptors in cutaneous T cell lymphoma. Sci Transl Med. 5:214ra1712013. View Article : Google Scholar
26	Torres-Cabala CA: Diagnosis of T-cell lymphoid proliferations of the skin: Putting all the pieces together. Mod Pathol. 33 (Suppl 1):83–95. 2019. View Article : Google Scholar
27	Cocks M, Porcu P, Wick MR and Gru AA: Recent advances in cutaneous T-cell lymphoma: Diagnostic and prognostic considerations. Surg Pathol Clin. 12:783–803. 2019. View Article : Google Scholar
28	Liebmann RD, Anderson B, McCarthy KP and Chow JW: The polymerase chain reaction in the diagnosis of early mycosis fungoides. J Pathol. 182:282–287. 1997. View Article : Google Scholar
29	Theodorou I, Delfau-Larue MH, Bigorgne C, Lahet C, Cochet G, Bagot M, Wechsler J and Farcet JP: Cutaneous T-cell infiltrates: Analysis of T-cell receptor gamma gene rearrangement by polymerase chain reaction and denaturing gradient gel electrophoresis. Blood. 86:305–310. 1995. View Article : Google Scholar
30	Wood GS, Tung RM, Haeffner AC, Crooks CF, Liao S, Orozco R, Veelken H, Kadin ME, Koh H, Heald P, et al: Detection of clonal T-cell receptor gamma gene rearrangements in early mycosis fungoides/Sezary syndrome by polymerase chain reaction and denaturing gradient gel electrophoresis (PCR/DGGE). J Invest Dermatol. 103:34–41. 1994. View Article : Google Scholar
31	Zelenetz AD: Guidelines for NHL: Updates to the management of diffuse large B-cell lymphoma and new guidelines for primary cutaneous CD30⁺ T-cell lymphoproliferative disorders and T-cell large granular lymphocytic leukemia. J Natl Compr Canc Netw. 12 (Suppl 5):797–800. 2014. View Article : Google Scholar
32	Liu WP, Song YQ, Zheng W, Wang XP, Ding N and Zhu J: Aggressive behavior and elevated lactate dehydrogenase at baseline confer inferior prognosis in patients with primary cutaneous lymphoma. Clin Lymphoma Myeloma Leuk. 13:534–540. 2013. View Article : Google Scholar
33	Quereux G, Frot AS, Brocard A, Leux C, Renaut JJ and Dreno B: Routine bone marrow biopsy in the initial evaluation of primary cutaneous B-cell lymphoma does not appear justified. Eur J Dermatol. 19:216–220. 2009. View Article : Google Scholar