Cold atmospheric plasma restores fulvestrant sensitivity by inhibiting CCND3 in the drug-resistant breast cancer cells – Nature

Executive Summary

This report details an investigation into the efficacy of Cold Atmospheric Plasma (CAP) in reversing drug resistance in fulvestrant-resistant breast cancer cells. The study aligns with the United Nations’ Sustainable Development Goal 3 (SDG 3), which aims to ensure good health and well-being, particularly by reducing premature mortality from non-communicable diseases like cancer. The findings demonstrate that CAP treatment effectively inhibits the growth of resistant breast cancer cell lines (T47D/fulR and MCF-7/fulR) and restores their sensitivity to fulvestrant. Genome-wide analysis identified the oncogene CCND3 as a key driver of resistance; its expression is elevated in resistant cells but significantly downregulated following CAP treatment. Functional validation confirmed that targeting CCND3 induces apoptosis and cell cycle arrest, mimicking the effects of CAP. This research presents CAP as an innovative therapeutic strategy (contributing to SDG 9: Industry, Innovation, and Infrastructure) with the potential to overcome endocrine therapy resistance, thereby advancing global efforts to improve cancer treatment outcomes and support the achievement of SDG 3.

1.0 Introduction: Aligning Cancer Research with Global Health Goals

1.1 The Global Burden of Breast Cancer and SDG 3

Breast cancer represents a significant global health challenge and a major impediment to achieving Sustainable Development Goal 3 (Good Health and Well-being), specifically Target 3.4, which calls for a one-third reduction in premature mortality from non-communicable diseases by 2030. Estrogen receptor-positive (ER+) breast cancer accounts for approximately 70% of cases. While endocrine therapies such as fulvestrant are effective, the development of drug resistance is a critical clinical problem leading to treatment failure and cancer recurrence. Overcoming this resistance is paramount for improving patient outcomes and making progress toward global health targets.

1.2 Innovative Therapeutic Approaches for Sustainable Health (SDG 9)

In line with SDG 9 (Industry, Innovation, and Infrastructure), which encourages scientific research and technological innovation, this study explores Cold Atmospheric Plasma (CAP) as a novel anti-cancer modality. CAP has shown promise in selectively targeting cancer cells. However, its potential to counteract resistance to specific drugs like fulvestrant remains largely unexplored. This report aims to elucidate the molecular mechanisms by which CAP may restore drug sensitivity in fulvestrant-resistant breast cancer cells, providing a potential new avenue for clinical application.

2.0 Report on Experimental Findings

2.1 CAP Restores Sensitivity to Fulvestrant in Resistant Cancer Cells

The study successfully established two fulvestrant-resistant (fulR) breast cancer cell lines, T47D/fulR and MCF-7/fulR, which required approximately 1,000-fold higher drug concentrations to inhibit growth compared to sensitive cells. The key findings on the effects of CAP were as follows:

• CAP treatment alone significantly inhibited the growth of both fulR cell lines.
• Pretreatment with CAP followed by fulvestrant administration resulted in a synergistic inhibition of cell growth, demonstrating a restoration of sensitivity to the drug.
• Colony formation assays and cell viability assays confirmed that CAP primes resistant cells to respond effectively to fulvestrant, an effect not observed in non-CAP-treated resistant cells.

2.2 CAP Modulates Key Genetic Pathways in Resistant Cells

To understand the molecular basis for this restored sensitivity, a genome-wide expression analysis was conducted. The analysis revealed that the acquisition of drug resistance and its reversal by CAP are associated with significant changes in critical cellular pathways.

• Gene Set Enrichment Analysis (GSEA) identified consistent dysregulation in genes related to ribosomal and mitochondrial protein-containing complexes during both the acquisition of resistance and its reversal by CAP.
• This suggests that CAP restores cellular sensitivity by normalizing fundamental processes of protein synthesis and energy metabolism, which are often hijacked by cancer cells to sustain resistance.

2.3 Identification and Validation of CCND3 as a Key Therapeutic Target

Among the numerous genes affected, the oncogene CCND3 was identified as a pivotal factor in the resistance mechanism.

1. Expression Pattern: CCND3 expression was significantly upregulated in both fulR cell lines. Following CAP treatment, its expression was markedly downregulated, suggesting it is a direct or indirect target of CAP therapy.
2. Functional Validation: To confirm the role of CCND3, it was silenced using siRNA in the resistant cell lines. This knockdown led to:
   • Decreased cell growth rates.
   • A synergistic increase in apoptosis (programmed cell death) when combined with fulvestrant treatment.
   • A significant extension of the G1 phase of the cell cycle, effectively arresting cell proliferation.

These results confirm that the downregulation of CCND3 is a key mechanism through which CAP restores fulvestrant sensitivity in resistant breast cancer cells.

3.0 Discussion: Implications for Sustainable Development

3.1 Advancing SDG 3: Good Health and Well-being

The findings presented in this report directly contribute to the objectives of SDG 3. By demonstrating a viable method to overcome endocrine therapy resistance, this research offers a pathway to improve the efficacy of existing cancer treatments. Re-sensitizing tumors to fulvestrant could extend patient survival, reduce cancer recurrence, and lower premature mortality from breast cancer, directly supporting the achievement of SDG Target 3.4.

3.2 Fostering SDG 9: Industry, Innovation, and Infrastructure

CAP represents a significant technological innovation in oncology. This study enhances scientific research (SDG Target 9.5) by uncovering the molecular mechanisms of a novel therapeutic technology. The successful application of plasma physics to solve a complex biological problem like drug resistance exemplifies the kind of cross-disciplinary innovation needed to address global challenges. Further development of CAP-based therapies could stimulate new industrial capabilities in medical technology.

3.3 Contribution to SDG 12: Responsible Consumption and Production

By restoring the effectiveness of an existing drug, CAP therapy promotes a more sustainable model of healthcare. It reduces the potential waste associated with ineffective treatments and may obviate the need for patients to move to more expensive and potentially more toxic second- or third-line therapies. This approach maximizes the utility of established medical resources, aligning with the principles of responsible consumption under SDG 12.

4.0 Conclusion

This study provides compelling evidence that Cold Atmospheric Plasma can reverse fulvestrant resistance in breast cancer cells by downregulating the oncogene CCND3 and modulating critical ribosomal and mitochondrial pathways. This innovative approach not only presents a promising new strategy for treating drug-resistant cancer but also aligns strongly with the global agenda for sustainable development. By advancing cancer treatment, fostering technological innovation, and promoting the efficient use of medical resources, this research contributes meaningfully to achieving SDG 3, SDG 9, and SDG 12, paving the way for more effective and sustainable cancer care worldwide.

Analysis of Sustainable Development Goals (SDGs) in the Article

1. Which SDGs are addressed or connected to the issues highlighted in the article?

The article primarily addresses two Sustainable Development Goals (SDGs):

• SDG 3: Good Health and Well-being: The entire study is focused on improving health outcomes for breast cancer patients. It investigates a novel therapeutic approach (Cold Atmospheric Plasma – CAP) to overcome drug resistance, a major challenge in cancer treatment. By aiming to restore the effectiveness of existing drugs like fulvestrant, the research directly contributes to the goal of ensuring healthy lives and promoting well-being.
• SDG 9: Industry, Innovation, and Infrastructure: The research represents a significant contribution to scientific innovation. The development and application of Cold Atmospheric Plasma (CAP) as a medical technology is an example of enhancing scientific research and upgrading technological capabilities. The study explores the molecular mechanisms of this innovative technology, aiming to pave the way for its clinical application, which aligns with fostering innovation.

2. What specific targets under those SDGs can be identified based on the article’s content?

Based on the article’s focus, the following specific targets can be identified:

1. Under SDG 3 (Good Health and Well-being):
   • Target 3.4: “By 2030, reduce by one third premature mortality from non-communicable diseases through prevention and treatment and promote mental health and well-being.” Breast cancer is a leading non-communicable disease (NCD). The research aims to develop more effective treatments for drug-resistant breast cancer, which directly contributes to reducing premature mortality from this disease. The article states, “overcoming fulvestrant resistance is of paramount importance for improving breast cancer treatment outcomes.”
   • Target 3.b: “Support the research and development of vaccines and medicines for the communicable and non-communicable diseases…” This study is a clear example of research and development for a new therapeutic approach (CAP) to treat an NCD (breast cancer). The article’s conclusion highlights its potential, stating it “pave[s] the way for further preclinical and clinical studies to validate CAP as a therapeutic approach for overcoming endocrine resistance.”
2. Under SDG 9 (Industry, Innovation, and Infrastructure):
   • Target 9.5: “Enhance scientific research, upgrade the technological capabilities of industrial sectors in all countries…including…encouraging innovation and substantially increasing the number of research and development workers…and public and private research and development spending.” The study is a direct output of enhanced scientific research, funded by the “National Research Foundation of Korea.” The authors from multiple research institutions are the R&D workers contributing to this target, and their work on an innovative technology like CAP exemplifies the goal of encouraging innovation.

3. Are there any indicators mentioned or implied in the article that can be used to measure progress towards the identified targets?

Yes, the article implies or directly mentions information related to several indicators:

1. For SDG 3 Targets:
   • Indicator 3.4.1 (Mortality rate attributed to…cancer…): While the article does not provide mortality statistics, its entire purpose is to develop a treatment that would ultimately lower the mortality rate for patients with drug-resistant breast cancer. The study’s findings on inhibiting cancer cell growth, increasing apoptosis, and restoring drug sensitivity are all proxy measures for progress that would contribute to reducing this indicator.
   • Indicator 3.b.2 (Total net official development assistance to medical research and basic health sectors): The article explicitly mentions its funding source: “This work was supported by the Basic Science Research Program (NRF-2022R1A2C1003483) of the National Research Foundation of Korea, funded by the Ministry of Education, Science and Technology.” This is a direct example of national investment in medical research, which is what this indicator measures.
2. For SDG 9 Targets:
   • Indicator 9.5.1 (Research and development expenditure as a proportion of GDP): The funding acknowledgment from the National Research Foundation of Korea is a concrete example of public R&D expenditure, which is the focus of this indicator.
   • Indicator 9.5.2 (Researchers…per million inhabitants): The list of authors and their affiliations (Department of Life Science, Dongguk University-Seoul; Plasma Bioscience Research Center, Kwangwoon University, etc.) represents the scientific workforce engaged in R&D, which this indicator aims to quantify.

4. Table of Findings

Source: nature.com