GUT MICROBIOTA EFFECTS IN HEMATOPOIETIC STEM CELL TRANSPLANT PATIENTS

Öz

Objective: In our study, we analyzed gut microbiota in allo-HSCT patients and aimed to evaluate the relationship of gut microbio- ta with transplant complications, mainly GVHD.
Materials and Methods: A total of 25 adult recipients and donors who underwent allo-HSCT at Istanbul Anadolu Medical Center were included in the study. Stool samples were collected twice, before chemotherapy regimen and after allo-HSCT. Samples were analyzed by High Melting (HRM) Analysis and Next Generation Sequencing (NGS) methods after nucleic acid isolation. Sequencing was done with Illumina MiSeq. Bacteria Silva database was used for taxonomic classification and QIIME 2 programs were used for analysis. Statistical analyses were carried out with the R statistical programming language.
Results: Twenty-five allo-HKHN recipients were included in the study. The mean age was 46.24±14.86 years in recipients and 43.40±13.20 years in donors. Gender distribution was M/F: 15/10 in patients and M/F: 17/8 in donors. Recipient and donor sibling HLA match was 10/10. The rate of GVHD associated with Allo-HSCT was 16%, and the relapse rate was 16%. It was observed that the Firmicutes and Proteobacteria phyla changed significantly before and after transplantation. The number of Entereccocus species was found to be higher in patients who developed GVHD and died. The loss of diversity was found to be statistically significant in the pre-transplant and post-engraftment samples of the patients. Conclusion: Gut microbiota diversity may guide the monitor- ing of GVHD and also may be manipulated for the treatment of GVHD. It is thought that increasing the diversity of commensal bacteria can also positively affect the prognosis of the disease.

Anahtar Kelimeler

• intestinal microbiota
• blood diseases
• hematopoietic stem cell transplantation
• HLA

ALLOJENİK KÖK HÜCRE NAKİLLERİNDE MİKROBİYOTA ETKİSİ

Öz

Amaç: Çalışmamızda allo-hematopoetik kök hücre nakli (al- lo-HKHN) uygulanmış hastaların mikrobiyota analizleri yapılmıştır. Nakile ve tedavilere bağlı olarak değişen mikrobiyota florasının engrafman ve Graft-Versus-Host Hastalığı (GVHH) gelişimi ile ilişkisinin gösterilmesi amaçlanmıştır.
Gereç ve Yöntem: İstanbul Anadolu Sağlık Merkezi’nde allo-HKHN uygulanan toplam 25 yetişkin alıcı ve vericileri çalışmaya dahil edildi. Dışkı örnekleri, HR öncesi ve allo-HKHN sonrası toplamda 2 kez alınmıştır. Örnekler, nükleik asit izolasyonu yapıldıktan sonra, Çözünürlüklü Erime Analizi (HRM) ve Yeni Nesil Dizileme (YND) yöntemi ile analiz edilmiştir. Dizileme işlemi, Illumina MiSeq cihazı ile yapılmıştır. Taksonomik sınıflandırma için Bacteria Silva veri bankası ve analiz için QIIME 2 programları kullanılmıştır. İstatistiksel analizler ise R istatistiksel programlama dili ile gerçekleştirilmiştir.
Bulgular: Çalışmaya dahil edilen alıcılarda yaş ortalaması 46,24±14,86 (18-71) yıl, vericilerde 43,40±13,20 yıl (11-61) olarak saptandı. Hastalarda cinsiyet dağılımı; E/K: 15/10 vericilerde E/K: 17/8 idi. Alıcı ve verici kardeş HLA uyumu 10/10 idi. Allo-HKHN ile ilişkili GVHH oranı %16, relaps oranı ise %16 bulundu. Nakil öncesi ve sonrası Firmicutes ve Proteobacteria filumlarının önemli ölçüde değiştiği gözlendi. GVHH geliştiren ve ex olan hastalarda Entereccocus türlerinin sayısı daha fazla bulundu. Hastaların nakil öncesi ve engrafman sonrası örneklerinde çeşitlilik kaybının istatistiksel olarak anlamlı olduğu saptandı.Sonuç: Allo-KHN sonrası artan patojen bakteriler ile hastalığın şiddetlenmesi, intestinal flora izlemi ile GVHH koruma ve/veya tedavisinde yönlendirici olabileceğini göstermektedir. Kom- mensal bakterilerin çeşitliliğinin arttırılmasıyla beraber hastalığın prognozunu da olumlu yönde etkileyebileceğini düşündürmektedir.

Anahtar Kelimeler

• Bağırsak mikrobiyotası
• kan hastalıkları
• hematopoetik kök hücre nakli
• HLA

Kaynakça

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