The impact of COVID-19 on microRNA and CD marker expression in AML patients | Scientific Reports – Nature.com

This study aimed to investigate the expression of microRNAs in leukaemia patients and their association with CD markers. Our results showed significant differences in the expression of microRNAs between AML patients and individuals in the control group. A strong correlation between microRNAs was observed, indicating that the presence of one microRNA led to increased expression of other microRNAs. Additionally, we found that MicroRNA16, MicroRNA21, and MicroRNA221 expression levels exhibited significant differences between AML patients with a history of COVID-19 infection and individuals without an infection history. Moreover, significant fold changes were observed between microRNA expression levels, and the CD markers among AML patients were positive for COVID-19. Finally, the upregulation of MicroRNA137 in AML patients was identified as a novel diagnostic biomarker that may aid in diagnosing, prognosis, and assessing treatment response for AML patients.

The diagnosis and classification of AML require a meticulous approach, as the World Health Organization (WHO) advised, encompassing a comprehensive blend of clinical history, morphological evaluation, cytogenetic/molecular genetic analysis, and immunophenotyping. Nonetheless, the absence of cytogenetic/molecular analyses in numerous healthcare facilities underscores the utility of cytomorphological evaluation supplemented by immunophenotyping, which proves adequate for expedited diagnosis and classification of AML (de Morais et al., 2022).

MicroRNAs are often deregulated in the cancerous process of body cells, and some of them enhance tumorigenesis and cancer progression by increasing tumour growth, angiogenesis, invasion, and immune evasion. The expression level of microRNAs in cancer can be used to predict patient prognosis and clinical response to treatment26,27. Since most of the microRNAs are intracellular, circulating microRNAs have been found in different body fluids and are identified as new cancer biomarkers. This study showed that the expression of microRNAs in AML patients increased significantly, and their role in the prognosis of AML disease was well established, similar to other studies28,29.

AML patients often have standard karyotypes30. They are a heterogeneous group from the molecular point of view, and these molecular differences are related to the prognosis of the disease. Therefore, it is crucial to identify new molecular markers and validate their promising future in treating the disease31. One of these essential markers is FLT3. It has been determined that it can be introduced as a valuable marker for diagnosing AML. In this study, we showed that the frequency of FLT3 mutation in AML patients was higher than in the control group and functions as an essential marker in the diagnosis and prognosis of AML disease; these results are consistent with the previous studies32,33,34,35. Fluorescence in situ hybridisation (FISH) diagnosis was used to detect chromosomal abnormalities in patients with AML, and based on the results of this study, FISH was able to show chromosomal abnormalities and disorders in patients with sufficient accuracy. Previous studies have investigated that the diagnostic strategies of FISH in diagnosing AML can be used as an efficient and powerful tool compared with other methods36. Additionally, FISH can be used as one of the standard methods for detecting microRNA expression, which was utilised in this study and effectively demonstrated the expression of microRNAs. Other studies have also shown the reliability of FISH as a method for detecting microRNA expression levels, which is consistent with the present studys findings. This technique has consistently proven its effectiveness in accurately assessing and quantifying the expression of microRNAs, confirming the results obtained in this research37,38.

Recent investigations have unveiled a variety of microRNAs that are dysregulated in AML and play roles in disease pathogenesis. Among these miRNAs are hsa-miR-16, hsa-miR-497, and hsa-miR-125, which have been implicated in regulating NRAS gene expression and cellular processes relevant to cancer development and progression (Hussen et al., 2021, Kipkeeva et al., 2022).

The BCR gene is a key player in the pathogenesis of several cancers, including AML. It encodes a protein regulating cell growth, differentiation, and survival. Dysregulation of the BCR gene, often through chromosomal translocations, can lead to the formation of oncogenic fusion proteins that drive leukemogenesis (Boucher et al., 2023). In the context of AML, aberrant expression of microRNAs has been implicated in disease progression. Specifically, miRNAs such as hsa-miR-16, hsa-miR-497, and hsa-miR-125 have been identified as potential regulators of gene expression and cellular processes relevant to cancer development. These miRNAs may target the BCR gene, influencing its expression levels and activity. Dysregulation of hsa-miR-16, hsa-miR-497, and hsa-miR-125 could thus contribute to aberrant BCR signaling and AML pathogenesis (Szczepanek, 2020). Understanding the regulatory interactions between genes and these miRNAs in AML patients is crucial for elucidating the molecular mechanisms underlying the disease and identifying potential therapeutic targets. Targeting the dysregulated expression of these miRNAs or modulating gene activity may represent promising strategies for AML treatment. Further investigation into the functional roles of hsa-miRNAs in gene regulation and AML pathogenesis is needed to validate their potential as therapeutic targets or diagnostic biomarkers.

Here, for the first time, we identified that the changes in the association between some of the microRNAs and CD1a, CD2, and CD11b were the most apparent alterations among AML patients after infection with COVID-19. Some of these findings are aligned with and confirmed by the previous study of a systematic screening of genetic events in AML patients, which shows that the expression of CD markers CD1a, CD2, and CD11b in AML patients is significantly increased39. The present study is also consistent with the analysis of Li et al.40, which shows increased expression of CD markers CD1a, CD2, and CD11b in Mixed-phenotype acute leukaemia (MPAL) patients, and they have revealed that these three CD markers are among the CD markers that have the highest expression and have the most changes.

The present study evaluated the levels of various cytokines in AML using flow cytometry for diagnostic purposes. Flow cytometric immunophenotypic analysis was conducted on all patients. The expression levels of CD99, CD33, CD45, CD13, HLA-DR, CD64, CD11c, and CD117 ranged from 54.58% to 90.56% in all cases. Meanwhile, MPO, CD38, CD36, CD15, CD34, CD11b, and CD2 exhibited expression levels between 24.48% and 45.06%, with the remaining CD markers showing expression levels below 20% in all cases. These findings were consistent with previous studies on AML (Rasheed et al., 2021, Piero et al., 2022). Previous researchers noted that markers such as MPO, CD13, CD33, CD15, and CD117 are specific to myeloid cells, while CD11c, CD64, CD14, and CD36 indicate monocytic lineage. Additionally, markers like CD19, CD22, CD10, CD79a, and CD20 are associated with B-lymphoid cells, while CD1a, CD2, CD3, CD4, CD5, CD7, and CD56 are linked to T-lymphoid/natural killer cells. Moreover, CD34, CD45, CD99, HLA-DR, Terminal deoxynucleotidyl transferase (TdT), and CD38 are commonly used as progenitor markers for diagnosing and classifying AML (Weir and Borowitz, 2001, Peters and Ansari, 2011).

Haematopoiesis is a highly regulated process controlled by complex molecular events that concurrently regulate hematopoietic stem cells commitment, differentiation, proliferation, and apoptosis. Substantial evidence now exists to demonstrate that microRNAs modulate haematopoiesis at the level of proliferation and differentiation and act as regulators of hematopoietic cell activity41. Differential expression of microRNAs in malignant cells compared to normal cells can be justified by their gene placement in cancer-related genomic regions, epigenetic mechanisms, and alterations in their processing mechanisms42,43. It has been established that variable expression of microRNAs plays a crucial role in leukemogenesis, and each cytogenetic alteration in AML is associated with the expression of a specific microRNA9,44. Generally, MicroRNA16 is a tumour suppressor that inhibits cell proliferation and induces apoptosis45. MicroRNA21 regulates immune responses in the body46, while MicroRNA125a, MicroRNA125b, and MicroRNA155 regulate hematopoietic stem cell activity8. MicroRNA192 plays a role in cell cycle arrest47, whereas MicroRNA221 induces cellular apoptosis48 and MicroRNA15a reduces apoptosis49. MicroRNA497 regulates cell proliferation50. In this study, the expression levels of these microRNAs were reported, and it was determined that they significantly impact disease progression. The increased expression levels of these microRNAs in AML patients are consistent with the findings of other studies that have reported higher expression of these microRNAs51,52. However, we presented for the first time the expression levels of microRNA and its correlation with CD markers in AML patients who were COVID-19 positive.

Considering that the expression patterns and levels of microRNAs can be used in disease prognosis and therapeutic responses23, the observed differences in the expression levels of microRNAs between AML patients and healthy individuals in this study are consistent with the results of other studies that have reported significant changes in the expression of microRNAs in various patients28,53. The decreased expression of microRNAs in the presence of CD markers and the concurrent decrease in CD marker expression in the presence of other CD markers align with the findings of studies by Shahrabi et al.11 and Gbarowska et al.54.

In a systematic review by Reyes-Long et al. (2023), they have demonstrated that microRNA expression levels differ between COVID-19 positive and negative patients, and these differences may be associated with the upregulation or downregulation of specific microRNAs55. Additionally, in the study by Shi et al. (2022), differences in microRNA expression among patients are observed56. The present study also found that the expression levels of microRNAs differed between COVID-19 positive and negative patients, and some microRNAs exhibited increased expression. The upregulation of microRNAs may not only be attributed to COVID-19 infection or vice versa but potentially due to the host's immune response to the virus57. Furthermore, considering the distinct expression levels of microRNAs in COVID-19 positive and negative patients, these changes in the expression levels of microRNAs could serve as suitable biomarkers58,59.

Identifying new types of microRNAs can be highly valuable and informative in understanding the mechanisms underlying various diseases, guiding therapeutic interventions, and promoting overall health improvement60,61. In the present study, microRNA 137 was identified as a novel biomarker involved in AML progression. MicroRNA137 plays a vital role in the evolution of the nervous system, the development of schizophrenia, and the maintenance of cellular homeostasis62,63. The present study showed an increased expression of MicroRNA137 in AML patients, while its expression was reduced in individuals who tested positive for COVID-19. However, a study by Wang et al. (2020) aimed to investigate the interaction between TRIM25 and MicroRNA137 and uncover their potential mechanisms in the progression of malignant AML. In that study, it was shown that there was a significant reduction in MicroRNA137 in blood samples of AML patients3. Cellular functional assays confirmed that the loss of MicroRNA137 substantially enhanced the invasion, migration, and proliferation abilities of AML cells. At the same time, overexpression of MicroRNA137 hindered the invasion, migration, and proliferation of AML cells. These findings indicated the tumour-suppressive role of MicroRNA137 in AML. However, further research is necessary due to the novelty of this microRNA. The precise significance and therapeutic target potential of MicroRNA137 may change as more is discovered about the molecular mechanisms behind AML and the involvement of microRNAs. Notably, research on MicroRNA137 and its role in AML is still underway. Targeting MicroRNA137in AML treatment may offer further therapeutic benefits, which will be further explored in subsequent research and clinical trials.

The present study demonstrated an increase in fold changes and microRNA expression in individuals who tested positive for COVID-19, and this increase was observed across all examined microRNAs. These findings are consistent with the results of studies conducted by Donyavi et al.59, Farr et al.64, and Li et al.65. This observation led us to understand that increased correlations for the expressions of genes in the cancer networks associated with decreased correlations for the expressions of genes in the standard networks might serve as valid biomarkers for early diagnosis of tumorigenesis and cancer progression. Finally, the identification of different MicroRNAs, the discovery of their target sequences, and the investigation of their gene expression pattern change in various cancers, including AML, along with their association with different cytogenetic abnormalities in AML patients, can provide a basis for more precise disease investigations, drug design, and the development of novel treatments for various types of cancer, including AML.

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The impact of COVID-19 on microRNA and CD marker expression in AML patients | Scientific Reports - Nature.com