Blood Res.  2021 Sep;56(3):156-165. 10.5045/br.2021.2020335.

The expression of multiple cancer/testis antigens can potentially be used to detect circulating disease and clonal evolution in the peripheral blood of multiple myeloma patients

• Shires K
• Wyk TV
• Wienand K

• KMID: 2520609
• DOI: http://doi.org/10.5045/br.2021.2020335

Abstract

Background
It is thought that cancer/testis antigens (CTAs) are expressed in a cascade-like manner in multiple myeloma as the disease progresses. In this pilot study, we investigated the co-expression of several CTAs in the peripheral blood (PB) during patient therapy to establish whether monitoring multiple CTAs allows for the prediction of relapse and clonal evolution.
Methods
We examined the co-expression of MAGEC1, MAGEA3, PRAME, and BAGE2 via quantitative reverse transcription-polymerase chain reaction (qRT-PCR) duplex assays in the PB mononuclear cells of 10 patients on chemotherapy at 3-month intervals, and correlated the levels to those of two basic clinical monitoring markers, serum β-2-microglobulin and serum M protein. Clonal evolution was investigated using flow cytometry to label the circulating malignant stem cell components with MAGEC1, PRAME, and MAGEA3 antibodies.
Results
Simultaneous monitoring of MAGEC1/PRAME provided sensitive detection of circulating malignant cells in easily accessible PB samples; transcript levels increased prior to changes in indicators of clinical relapse. While MAGEA3/BAGE2 expression levels did not offer earlier prediction of relapse, they provided insight into significant changes occurring within the malignant cell population; the addition of either CTA to a MAGEC1-monitoring panel allowed for better classification of the relapse event (clonal evolution), which in turn could potentially guide treatment strategies in the future.
Conclusion
This pilot study supports the novel idea of determining the levels and CTA expression patterns of the total circulating malignant cell population (pro-B/pre-B stem cell progenitors and proliferating plasma cells) as an alternate disease monitoring methodology.

Keyword

CTA; MAGEC1; Myeloma; Cascade; Monitoring; PRAME

Figure

• 

  Fig. 1 PRAME transcript expression in MM patients during chemotherapy. RNA samples collected from patients MM1, 4, and 6 at diagnosis and during treatment were analyzed for PRAME expression using the PRAME/ABL qRT-PCR assay. The normalized ratios were plotted along with the previously collected normalized MAGEC1 data [21] and serum M and Sb2M levels. *Indicates patient death within three months of the last data point; C+D, cyclophosphamide/dexamethasone cycles; M+P, melphalan/prednisone cycles; R, localiszd radiation; open circles indicate a lack of detectable transcripts at a sensitivity level determined by the corresponding ABL Cq.

• 

  Fig. 2 MAGEA3 transcript expression in MM patients during chemotherapy. RNA samples collected from patients MM 3, 8, and 10 at diagnosis and during treatment were analyzed for MAGEA3 expression using the MAGEA3/ABL qRT-PCR assay. The normalized ratios were plotted along with the previously collected normalized PRAME data, MAGEC1 data [21], and serum M levels. *Indicates patient death within three months of the last data point; C+D, cyclophosphamide/dexamethasone cycles; R, localized radiation. Open circles indicate a lack of detectable transcripts.

• 

  Fig. 3 BAGE2 transcript expression in MM patients during chemotherapy. RNA samples collected from patients MM 6, 8, and 9 at diagnosis and during treatment were analyzed for BAGE2 expression using the BAGE2/ABL qRT-PCR assay. The normalized ratios were plotted along with the previously collected normalized MAGEA3, PRAME, and MAGEC1 data and serum M/Sβ2M levels. *Indicates patient death within three months of the last data point; colored arrows indicate the date of associated transcript increase, showing a succession pattern; Crt, creatinine mmol/l; Hb, hemoglobin g/dL; open circles indicate a lack of detectable transcript.

• 

  Fig. 4 Schematic of the proposed evolution of MM clones based on the expression of a CTA cascade.

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