Today, we are pleased to introduce Dr. Alfonso Calvo, a principal investigator at Cancer Center Clínica Universidad de Navarra (CCUN) in Spain. His research focuses on understanding how changes in the tumor microenvironment (TME) contribute to resistance against immunotherapies in non-small cell lung cancer (NSCLC).
When his team faced a critical technical challenge engineering a lung cancer cell line, Dr. Calvo turned to Cytosurge’s CellEDIT service for support. Our collaboration helped advance his research, bringing him one step closer to important new insights in cancer immunotherapy.

Alfonso Calvo González, Professor of Cell Biology and Histology, School of Medicine and Sciences, University of Navarra, Pamplona, Spain. Director of the Department of Pathology, Anatomy and Physiology, School of Medicine, University of Navarra, Pamplona, Spain. Researcher (PI), Program of Solid Tumors, CIMA of the University of Navarra, Pamplona, Spain.
Tell us something about your project.
Our project focuses on understanding how PTEN loss alters the tumor microenvironment (TME) and leads to resistance to immunotherapy, specifically anti-PD1 treatment. In particular, PTEN is a tumor suppressor gene that plays a key role in controlling cell growth and survival. Loss of PTEN function can disrupt these processes, contributing to an immunosuppressive tumor microenvironment and resistance to therapies like anti-PD1 treatment.
We had already demonstrated these effects in a squamous carcinoma model (Cancer Res (2023) 83 (15): 2513–2526). To validate our findings that PTEN loss contributes to resistance against anti-PD1 therapy by altering the tumor microenvironment, we aimed to create a PTEN knockout. The Lacun.3 lung adenocarcinoma cell line was chosen for its aggressiveness, making it an ideal model for our study.
What challenge were you facing in your project?
Despite applying protocols that had worked successfully in the past in our squamous carcinoma model, we couldn't achieve the knockout in Lacun.3. After several unsuccessful internal attempts, we realized we needed external expertise to move forward.
How did you discover CellEDIT, and what convinced you to work with us?
I found CellEDIT while researching online. I didn’t know about Cytosurge or the technology initially, but after speaking with Dr. Gorka Santos and other team members, I felt confident. What convinced me was the combination of competitive pricing, a manageable timeline, and clear, professional communication.
Cytosurge uses a unique methodology involving single-cell nuclear injection of editing components with FluidFM technology. This technique was different from our approach and any other method I knew about. However, since our own strategies were not yielding the desired results, I trusted their expertise. In the end, they achieved the knockout we needed.
How would you describe your experience working with CellEDIT?
The collaboration with the CellEDIT team was very smooth. The communication was excellent; the team regularly updated us on the project’s progress. Even though it was my first time outsourcing such a service, the workflow was easy to follow. Their scientific knowledge, responsiveness, and ability to adapt the methodology to our specific needs made a big difference.
How important was CellEDIT's support for your research progress?
Crucial. Without the successful PTEN knockout, we wouldn’t be able to complete our planned experiments or validate our hypotheses. Thanks to CellEDIT’s work, we are now finalizing the study and hope to submit our results for publication by the end of the year.
Would you work with CellEDIT again?
Yes, most probably, especially for similarly complex projects. Of course, budget constraints are always a factor in academic research, but if outsourcing is necessary, CellEDIT would be my first choice based on this positive experience.
Dr. Alfonso Calvo’s story highlights how tailored solutions, strong communication, and expert methodologies can transform research obstacles into important scientific milestones. We are honored to have contributed to advancing his project and look forward to the impactful discoveries to come.
Turning CRISPR Challenges into Breakthroughs: How CellEDIT Empowered Cancer Research at the University of Navarra
A Conversation with Dr. Alfonso Calvo