pattern
pattern

Background

What is Obesity and Why is it a Problem

Obesity is a complex medical condition characterized by an excessive amount of body fat.[1]

It is clinically defined by a Body Mass Index (BMI) of 30 or higher.[2]

This condition has escalated into a global pandemic, affecting over a billion people worldwide and acting as a primary driver for numerous chronic diseases.[3]

The health consequences are severe, as obesity significantly increases the risk of developing life-threatening conditions such as type 2 diabetes, cardiovascular diseases (including heart attacks and strokes), hypertension, and several types of cancer.[4]

This not only diminishes the quality of life for individuals but also places an immense and growing strain on global healthcare systems.[5]

Growing Obesity Issues

Figure 1. Growing Obesity Issues

History of Obesity Treatments

The approach to treating obesity has evolved significantly over the decades.[6]

Early strategies focused almost exclusively on diet and exercise, but these methods often yielded unsustainable results for many individuals.[7]

The 20th century saw the introduction of pharmaceutical interventions, beginning with appetite suppressants that often had dangerous side effects.[8]

In recent years, a new class of drugs, GLP-1 receptor agonists like Wegovy (semaglutide), has shown remarkable efficacy in weight reduction.[9]

However, their high cost, significant gastrointestinal side effects, and the common issue of weight regain after discontinuation highlight the limitations of current pharmacological approaches and underscore the urgent need for safer, more accessible, and sustainable therapeutic alternatives.[10]

History of Obesity treatment

Figure 2. History of Obesity treatment

What is the Microbiome?

The human microbiome refers to the vast and diverse community of trillions of microorganisms—including bacteria, viruses, fungi, and archaea—that reside in and on our bodies.[11]

The gut microbiome, located in the gastrointestinal tract, is the most densely populated and metabolically active of these communities.[12]

Far from being passive inhabitants, these microbes play a critical and symbiotic role in human health, influencing everything from digestion and nutrient absorption to immune system development and the production of essential vitamins.[13]

Human Microbiome

Figure 3. Human Microbiome

The Microbiome and Obesity Relationship

A growing body of scientific evidence reveals a strong and intricate relationship between the gut microbiome and metabolic health, including obesity.[14]

The composition of gut bacteria in individuals with obesity is often significantly different from that of lean individuals, frequently showing an altered ratio of major bacterial phyla like Firmicutes and Bacteroidetes.[15]

These microbes influence weight regulation through several mechanisms: they help harvest energy from the diet, produce key metabolites like short-chain fatty acids (SCFAs) that regulate energy metabolism, and modulate host immune responses and intestinal barrier function. [16]

As shown in preclinical studies, specific probiotic species, such as Lactobacillus, have been demonstrated to have a direct impact on weight regulation and lipid metabolism in mouse models, suggesting that modulating the microbiome could be a powerful therapeutic strategy against obesity.[17]

Gut Microbiome and Weight regulation

Figure 4. Gut Microbiome and Weight regulation

Molecular Network of Adipogenesis and AMPK

Our project targets the fundamental cellular process of fat creation, known as adipogenesis, which is controlled by a complex molecular network.[18]

Molecular intervention part 1

Figure 5. Adipogenesis and its Chemical Induction (DMI)

Adipogenesis and its Chemical Induction (DMI): Adipogenesis is the process by which pre-adipocyte cells differentiate into mature, lipid-storing adipocytes (fat cells).[19] In the laboratory, this process can be reliably induced in cell lines like 3T3-L1 using a chemical cocktail known as DMI.[20] This cocktail consists of Dexamethasone (a glucocorticoid), IBMX (a phosphodiesterase inhibitor), and Insulin. Each component activates distinct signaling cascades that converge to initiate the adipogenic program.[21]

Molecular intervention part 2

Figure 6. Key Adipogenesis Genes (PPARγ and C/EBPα)

Key Adipogenesis Genes (PPARγ and C/EBPα): The differentiation process is governed by a cascade of transcription factors.[22] The two master regulators at the heart of this network are PPARγ (Peroxisome proliferator-activated receptor gamma) and C/EBPα (CCAAT/enhancer-binding protein alpha).[23] These proteins are essential for the expression of genes involved in creating and storing lipids, and their activation is a critical step for a cell to become a mature adipocyte. Therefore, inhibiting these master regulators is a primary strategy for preventing fat accumulation.[24]

Molecular intervention part 3

Figure 7. AMPK: The Master Metabolic Regulator

AMPK: The Master Metabolic Regulator: AMP-activated protein kinase (AMPK) is a crucial cellular energy sensor that acts as a master metabolic switch.[25] When cellular energy levels are low, AMPK is activated, and it works to restore energy balance by shutting down energy-consuming processes (like fat synthesis, or lipogenesis) and ramping up energy-producing processes.[26] Critically, the activation of AMPK has an inhibitory effect on adipogenesis. It directly suppresses the expression and activity of the master regulators PPARγ and C/EBPα.[27] This makes AMPK a prime therapeutic target: by activating AMPK, we can effectively turn off the molecular machinery responsible for creating new fat cells.[28]

Adipogenesis Schematic Protocol

Adipogenesis Schematic Protocol

Figure 8. 3T3-L1 cells are seeded into the plate (D-2) and wait until full confluency(D-0). BCS media is changed into DMI media(D-0) in 48 to72 hours (D-3). Media is changed into FI media for 48hours(D-5) after that change normal FBS media for 48hours(D-7).

References

[1] Taieb, A. B., et al. (2022). Understanding the risk of developing weight-related complications. Diabetology & Metabolic Syndrome.
[2] Centers for Disease Control and Prevention. (2024). Obesity and Severe Obesity Prevalence in Adults (BMI ≥30).
[3] Chen, E. (2024, February). More than a billion people have obesity, study estimates. Stat News.
[4] Islam, A. N. M. S., et al. (2024). The global burden of overweight–obesity and its associated health risks. BMC Public Health.
[5] Yu, C. Y., et al. (2014). Statistical methods for body mass index: a selective review of the literature. arXiv preprint.
[6] Bray, G. A. (2022). An Historical Review of Steps and Missteps in the Management of Obesity. In Obesity: Prevalence, Mechanisms, and Management (pp. 1–20).
[7] Haganes, K. L., et al. (2025). Maintenance of time-restricted eating and high-intensity interval training for weight management. Scientific Reports.
[8] Dolgin, E. (2012). A history of anti-obesity medications in the United States. Nature Medicine.
[9] Scragg, J. (2025). The societal implications of using glucagon-like peptide-1 receptor agonists for weight loss. Obesity Reviews.
[10] Quarenghi, M., et al. (2025). Weight Regain After Liraglutide, Semaglutide or Placebo. Journal of Clinical Medicine.
[11] Lloyd-Price, J., Abu-Ali, G., & Huttenhower, C. (2016). The healthy human microbiome. Genome Medicine.
[12] Hou, K., et al. (2022). Microbiota in health and diseases. Signal Transduction and Targeted Therapy.
[13] Oliphant, K., & Allen-Vercoe, E. (2019). Macronutrient metabolism by the human gut microbiome. Microbiome.
[14] Turnbaugh, P. J., et al. (2006). An obesity-associated gut microbiome with increased capacity for energy harvest. Nature, 444(7122), 1027–1031.
[15] Ley, R. E., Turnbaugh, P. J., Klein, S., & Gordon, J. I. (2006). Microbial ecology: Human gut microbes associated with obesity. Nature, 444(7122), 1022–1023.
[16] Cani, P. D., & Delzenne, N. M. (2009). The role of the gut microbiota in energy metabolism and metabolic disease. Current Pharmaceutical Design, 15(13), 1546–1558.
[17] Park, D. Y., et al. (2013). Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032 cell extracts exert anti-obesity effects in high-fat diet-induced obese mice. Nutrition & Metabolism, 10(1).
[18] Rosen, E. D., & Spiegelman, B. M. (2000). Molecular regulation of adipogenesis. Annual Review of Cell and Developmental Biology, 16, 145–171.
[19] Green, H., & Kehinde, O. (1975). An established preadipose cell line and its differentiation in culture. Cell, 5(1), 19–27.
[20] Student, A. K., Hsu, R. Y., & Lane, M. D. (1980). Induction of fatty acid synthetase synthesis in differentiating 3T3-L1 preadipocytes. Journal of Biological Chemistry.
[21] Zebisch, K., Voigt, V., Wabitsch, M., & Brandsch, M. (2012). Protocol for effective differentiation of 3T3-L1 cells to adipocytes. Analytical Biochemistry, 425(1), 88–90.
[22] Farmer, S. R. (2006). Transcriptional control of adipocyte formation. Cell Metabolism, 4(4), 263–273.
[23] Tontonoz, P., & Spiegelman, B. M. (2008). Fat and beyond: the diverse biology of PPARγ. Annual Review of Biochemistry, 77, 289–312.
[24] Rosen, E. D., et al. (2002). C/EBPα induces adipogenesis through PPARγ: a unified pathway. Proceedings of the National Academy of Sciences, 99(14), 10488–10493.
[25] Hardie, D. G. (2011). AMP-activated protein kinase: an energy sensor that regulates all aspects of cell function. Genes & Development, 25(18), 1895–1908.
[26] Carling, D., et al. (2008). AMP-activated protein kinase: balancing the scales. Biochimie, 90(1), 9–18.
[27] Habinowski, S. A., & Witters, L. A. (2001). The effects of AICAR on adipocyte differentiation of 3T3-L1 cells. Biochemical and Biophysical Research Communications, 286(5), 852–856.
[28] Giri, S., et al. (2006). AMP-activated protein kinase (AMPK) inhibits adipogenesis through modulation of mitotic clonal expansion and C/EBPα function. Biochemical and Biophysical Research Communications, 349(1), 91–98.