ABOUT POLYCYSTIC OVARIAN SYNDROME

PCOS is a metabolic syndrome of unknown aetiology with inconsistent symptoms. Recent research suggests its plausible cause to a combination of genetic factors and lifestyle changes. PCOS is manifested through various hormonal imbalances in susceptible ovary owners. This may result in an unnatural enlargement of the ovaries and/or formation of multiple cysts. Other related problems could be severe metabolic disorders such as Type 2 diabetes, cardiovascular diseases or some cancers in a few extreme cases.

As per World Health Organization (WHO), it is estimated that 116 million women (3.4%) are affected by PCOS worldwide, with its prevalence in our country being estimated to range from 3.7 to 22.5 per cent, i.e. 2 out of 10 women in India suffer from PCOS [1,2].

ASSOCIATED SYMPTOMS

As PCOS is a syndrome, it is a set of numerous symptoms. These symptoms can range from non-hampering to severe metabolic disorders affecting the patient for a lifetime [3-5]. Depending on the method of diagnosis, the prevalence of PCOS cases varies significantly from 6% to 12%, making it the most common female endocrinopathy disorder during reproductive life [6].

Some of these symptoms include:

MANAGEMENT OF SYMPTOMS AND CURRENT DIAGNOSTIC

PCOS diagnosis has always proven to be problematic. There exists various criteria for diagnosing a person with PCOS, like the Rotterdam criteria, NIH Criteria, and AES criteria (Click Background to know more about criteria). However the existing diagnosis methods cannot be used to their potential due to lack of basic healthcare centres, trained medical professionals, expensive ultrasound machines and blood test kits. During our analysis we found that many government healthcare centres lack basic amenities, which reduces the accessibility to such tests. Laboratories across various states face problems of poor infrastructure, ill-equipped one-room labs in public clinics, and small private neighbourhood labs with limited testing equipment. Additionally, the cost of having these tests exceeds the average annual income of families with income below the poverty line. This poses a threat to the diagnosis of the syndrome. The absence of an accurate and reliable diagnosis method prevents opportunities for various studies for PCOS, which affects research for this syndrome. Due to this, finding its aetiology becomes more complex, further slowing the development of promising treatments. To add to the problem, the system still relies heavily on patients’ initiative and awareness to ensure successful testing. Tackling the diagnosis issue becomes crucial to the management and finding a cure for PCOS. Thus, a kit that is easier to use and accessible for all, irrespective of their socio-economic background, is the need of the hour [7,8].

APTASTELES

A novel diagnostic kit for PCOS

Realising the importance of a diagnostic kit for PCOS, our team started working upon making it a reality. We came up with ‘AptaSteles’, a novel diagnostic kit for PCOS. 

The kit diagnoses the syndrome based on a range of biomarkers that accouts for many of the symptoms(Click Background to know more about biomarkers). After an intensive literature review, our team focused on the following biomarkers:

For the detection of these biomarkers, we have harnessed the power of aptamers (short oligonuleotide sequence that bind specifically to molecules by the formation of particular secondary structures). A class of aptamers called light up aptamers (aptamers which bind to molecules called fluorophore and give fluorescent output) is used for the visualisation of the interaction between aptamer - biomarker and quantification of the biomarkers. Levels of these biomarkers are detected, and their combination is analysed to give a quantitative result for the presence of PCOS (Click Background to know more about aptamers and light up aptamers).

The proposed kit is made up of the following components:

1. Hormone and Protein Detection: We incorporated a dual aptamer approach consisting of two aptamers - recognition module and amplification module. The recognition module consists of a biomarker specific aptamer fused with additional sequences called trigger. The amplification module is a target sequence which consists the complementary sequence to trigger, promoter and the DNA sequence required for production the light up aptamer. 



Figure 5. Recognition and amplification modules



2. miRNA Detection: The FASTmiR (Fluorescent Aptamer Sensor for Tracking miRNAs) approach is used to detect miRNAs. It is an RNA sequence of the complementary sequence to the miRNA of interest and a light up aptamer.



Figure 6.FASTmiR

We synthesised a DNA library for two of our biomarkers that do not have characterised aptamers and plan to perform SELEX (Systematic Evolution of Ligands by Exponential Enrichment)(Click Background to know more about SELEX).

An essential element of the kit is the amplification of miRNA using a technique called microRNA Recombinase Polymerase Amplification (miRPA), to accommodate the low concentrations of biomarkers that might be present in the blood/plasma of the users.

These detection modules are integrated into a point of care device. The device is built on the principles of microfluidics, complemented by the optical detection components required by the hardware device. 

(Click Blueprint to get a better idea about the approaches and how we are using them to diagnose PCOS).

The aptamer-based detections for biomarkers, SELEX, and our kit hardware design all are approaches to make AptaSteles an all-inclusive project to shape our proposed kit. We hope these baby steps will lead us to make a diagnostic kit that is safe, accurate, and user-minded.

REFERENCES

1. Ganie, M. A., Vasudevan, V., Wani, I. A., Baba, M. S., Arif, T., & Rashid, A. (2019). Epidemiology, pathogenesis, genetics & management of polycystic ovary syndrome in India. The Indian journal of medical research, 150(4), 333.
2. Bulsara, J., Patel, P., Soni, A., & Acharya, S. (2021). A review: Brief insight into Polycystic Ovarian syndrome. Endocrine and Metabolic Science, 3, 100085.
3. https://www.nhs.uk/conditions/polycystic-ovary-syndrome-pcos/symptoms/
4. https://www.hopkinsmedicine.org/health/conditions-and-diseases/polycystic-ovary-syndrome-pcos
5. https://www.mayoclinic.org/diseases-conditions/pcos/symptoms-causes/syc-20353439
6. Palomba, S., Santagni, S., Falbo, A., & La Sala, G. B. (2015). Complications and challenges associated with polycystic ovary syndrome: current perspectives. International journal of women's health, 7, 745.
7. Engel, N., Ganesh, G., Patil, M., Yellappa, V., Pant Pai, N., Vadnais, C., & Pai, M. (2015). Barriers to point-of-care testing in India: results from qualitative research across different settings, users and major diseases. PLoS One, 10(8), e0135112.
8. Wolf, W. M., Wattick, R. A., Kinkade, O. N., & Olfert, M. D. (2018). Geographical prevalence of polycystic ovary syndrome as determined by region and race/ethnicity. International journal of environmental research and public health, 15(11), 2589.