Mikrochimérismus – na hraně počínajícího relapsu AML?

Znovuobjevení či nárůst autologní krvetvorby po alogenní transplantaci hematopoetických buněk bývá spojen s počínajícím relapsem akutní myeloidní leukemie, nicméně význam mikrochimérismu (podílu autologních buněk pod hranicí 1 %) není jednoznačný. Byly porovnávány výsledky fragmentační analýzy (FA) a kvantitativní PCR v reálném čase (RQ-PCR). Hodnoty získané z obou analýz byly konfrontovány s klinickým stavem pacientů: kompletní remise, molekulární relaps, hematologický relaps. Zatímco RQ-PCR analýza periferní krve predikovala hematologický relaps ve 12 případech z 13 (92 %), s mediánem předpovědi 34 dnů (0–322 dní), byla FA úspěšná pouze v pěti případech (38 %) (medián předpovědi 0 dní, 0–154 dní) (p = 0,002). RQ-PCR předcházela FA v 10 případech, s mediánem dřívější předpovědi 35 dní (interval 6–168 dní). RQ-PCR dále zachytila dalších 4 z 6 časných hematologických relapsů (do jednoho roku po transplantaci), mimo dosah senzitivity FA. Kombinace vyšetření periferní krve a kostní dřeně navíc potvrdilo 9 molekulárních relapsů z 9. RQ-PCR se jeví jako citlivá a spolehlivá metoda stanovení mikrochimérismu, jehož dynamika umožňuje zachycení počátku relapsu a umožňuje tak časnou klinickou intervenci.

Klíčová slova:
chimérismus, mikrochimérismus, relaps, AML, RQ-PCR

Microchimerism – on the edge of incipient AML relapse?

The reoccurrence or increase in autologous hematopoiesis after allogeneic stem cell transplantation has been linked with incipient leukemia relapse, however, importance of such emergency regarding microchimerism (chimerism under 1% of autologous cells) remains controversial. We compared conventional PCR of minisatellite, microsatellite, or sex specific regions followed by fragment analysis (FA) with real-time quantitative PCR (RQ-PCR) of insertion/deletion and sex polymorphism. Chimerism values obtained from both analyses were compared with the status of the disease (continuous complete remission, molecular relapse, hematological relapse, HR). Whereas RQ-PCR analysis of microchimerism predicted full HR in 12/13 cases (92%) with median of 34 days (0-322 days), the conventional FA was successful only in 5/13 cases (38%) (median 0 days, interval 0-154 days; p = 0.002). RQ-PCR surpassed FA in 10 cases with median of earlier prediction of relapse of 35 days (interval 6-168 days). Besides this, RQ-PCR detected other 4/6 HR until one year after transplantation, beyond the sensitivity of FA. Further, RQ-PCR examination of peripheral blood in combination with bone marrow confirmed all 9 molecular relapses previously detected due to a specific molecular marker. We conclude, that RQ-PCR is sensitive and reliable method of microchimerism analysis that can predict impending relapse and allows therefore earlier clinical intervention in comparison with another, less sensitive method.

Key words:
chimerism, microchimerism, relapse, AML, RQ-PCR

Autoři: O. Horký; J. Mayer; L. Kablásková; M. Borský; M. Krejčí; D. Dvořáková
Působiště autorů: Centrum molekulární biologie a genové terapie, Interní hematoonkologická klinika LF MU a FN Brno
Vyšlo v časopise: Transfuze Hematol. dnes,15, 2009, No. 2, p. 97-102.
Kategorie: Souhrnné práce, původní práce, kazuistiky

Souhrn

Klíčová slova:
chimérismus, mikrochimérismus, relaps, AML, RQ-PCR

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