Company: APM
Filing Date: 2025-12-05
Form Type: 424B5
Source: 0001213900-25-118752
Chunk: 292

Company: Aptorum Group Ltd
Filing Date: 2025-12-05
Form: 424B5
Chunk 292
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 miRNAs correlate with a disease, and each method has its limitation: first, expression patterns
of hundreds of miRNAs in a bodily fluid from patients with a pathology of interest and from control subjects are compared using RT-qPCR,
miRNA array or next generation sequencing (NGS). NGS is a technology for DNA and RNA sequencing and variant/mutation detection. This
technology combines the advantages of unique sequencing chemistries, different sequencing matrices, and bioinformatics technology. Such
a combination allows a massive parallel sequencing of various lengths of DNA or RNA sequences or even whole genome within a relatively
short period of time. NGS involves several major steps in sequencing: DNA fragmentation, library preparation, parallel sequencing, and
bioinformatics analysis, and variant/mutation annotation and interpretation.

Second, analysis of disease-specific
miRNAs is performed by comparing miRNAs isolated from pathologic and normal tissue, organ, or cells (MicroRNA profiling: approaches and
considerations. Colin C. Pritchard et. al., Nature Reviews Genetics volume 13, pages 358 – 369 (2012)).
Both approaches hold promise; however, because the identified biomarkers are not restricted to a particular organ or tissue, they are
often not sufficiently sensitive and/or specific to be of practical use. (Advances in multiplexed techniques for the detection and quantification
of microRNAs. Jet et. al., Chem. Soc. Rev., 2021, 50, 4141-416 &Direct detection and quantification of microRNAs. Hunt
et. al. Anal Biochem. 2009 Apr 1; 387(1): 1 – 12).

<div align='center'>164

Summary of Current Approaches to Analysis of miRNA in Plasma</div>

In the table above, the main
advantage of the first approach [miRNA Array/NGS], i.e. the ability to test hundreds of miRNAs and its disadvantages, namely lower
sensitivity and higher variability as compared to RT-PCR are reported in the following references.

Williams Z, Ben-Dov IZ, Elias
R, Mihailovic A, Brown M, Rosenwaks Z, Tuschl T. Comprehensive profiling of circulating microRNA via small RNA sequencing of cDNA
libraries reveals biomarker potential and limitations. Proc Natl Acad Sci U S A. 110, 4255-4260 (2013). PMID: 23440203.

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