1 INTRODUCTION

Priscille did her PhD at the Institut Gustave Roussy on the pharmacology of HIV integrase and then continued her Post‐Doctoral research at the Institut Pasteur, Paris, France with Professor Stewart T. Cole. After becoming a fellow from INSERM, Priscille moved to the Institut Pasteur Korea, Seoul in 2006 and started her team with the support of the Avenir Program. In 2011, Priscille was laureate of an ERC‐ Consolidator grant and relocated her team in Lille, France at the crossroads of Europe. She is interested in how pathogenic mycobacteria interact with their host cells and how small molecules inhibitors can thwart these interactions. She has developed a passion for high content screening and enjoys sharing her technical expertise to solve key questions on host‐pathogens interactions.

1.1 What is your research background ‐ what did you first start working on at the beginning of your academic career?

I studied biology at the chemical engineering school of Montpellier, France. I did two trainings during my Master 2, the first one on the delivery of oligonucleodides by the HIV TAT peptide and the second one on two‐hybrid approaches on the male determination factor SRY. Straight after that, I moved to Paris for my PhD on the development of inhibitors of HIV integrase at the Institut Gustave Roussy. It was early 1997, when the multi‐therapy combining anti‐reverse transcriptase and anti‐protease inhibitors had just been launched for HIV treatment and appeared very promising. So targeting the third HIV enzyme seemed the way to go. The initial objective was to make a DNA triplex between the retro‐transcribed viral genome and an oligonucleotide to prevent the binding of the integrase to its substrate and I ended up with the selection of an aptamer!

1.2 Have you always wanted to work in academia?

Actually no. After my PhD, I hesitated to join a biotech company. In addition, I also had hesitations between opportunities in North America or in France. However, when I was offered to join the Institut Pasteur on a tuberculosis (TB) vaccine project based on comparative genomics, I was thrilled and did not hesitate to join the lab. of Stewart Cole. Actually, I really enjoyed my post‐doctoral time and often say to my new students that the post‐doc period is the best period ever in a research career. The Institut Pasteur is really an exciting place for many research themes and by joining an EU project, I had the opportunity to establish very nice and fruitful connections with different labs in the EU. During that time, I applied for tenure track positions in French Academia and was successful at INSERM in 2005, so I went for it.

1.3 How has your early work brought you to what you are working on now?

I always enjoyed working on microbes and designing strategies to combat them! My current research is mostly on Mycobacterium tuberculosis (Mtb), with which I have become familiar during my post‐doc. Roland Brosch and Alex Pym taught me all the basics. Then, I was offered an exciting research opportunity in the newly created Institut Pasteur Korea, an all‐new research environment including the building design process. There, I managed to develop phenotypic assays on virulent Mtb strains suitable for the high throughput screening of libraries of modulators. These approaches undertaken together with Thierry Christophe turned out to be very successful, with for instance the discovery of Q203, currently a Phase IIB drug candidate. Back in France, I pursued the quest for new drugs with research projects focused on deciphering novel mechanisms of action. We have further extended these approaches to other bacteria and microbes. Notably, we set‐up and performed image‐based phenotypic screenings on SARS‐Cov2 infected cells during lock‐down.

1.4 Have you always been interested in cellular microbiology?

I have always been interested in chemistry‐microbiology. I was thrilled by the exploration of various paths in omics. Thus, the implementation of comparative and functional genomics in my research, pioneering at the time, led to the discovery of the type 7 secretion system in Mtb. I still vividly remember my astonishment when I saw the phenotype of the first SCID mice infected with BCG overexpressing RD1 (BCG‐RD1)! Then, we screened more than 200,000 compounds over a 3‐year period, which resulted in the development of Q203. This was certainly a very privileged moment in my career, with the crystallisation of the desire to contribute to the discovery of anti‐TB drugs. This was yet another exciting moment, with evidence for the in vivo efficacy of the new drug candidate! Then, I was awarded an ERC to study fundamental questions, in particular host‐directed approaches against Mtb, which were considered to be pioneer research in 2010, but has now become very popular in the microbiology field.

1.5 What do you think the most interesting questions are at the moment in cellular microbiology?

First, TB treatments rely on the taking of a combination of drugs for several months. Usually four drugs. Very effective, but slow acting and subject to resistance. How to keep this combo efficient and avoid the appearance of drug resistance and tolerance? I believe that a better understanding of the host response following the administration of these antibiotics is part of the key.

Second, the interaction of Mtb with various cells in the lungs and in the body has also be curtailed by the importance given to the Macrophage niche. The local lung epithelium or the sensory neurons signals could also contribute to the macrophage response and this deserves to be explored with the newly developed tools such as organoids or lung‐on‐chip.

Third, Mtb uses also numerous strategies to successfully complete the various steps of infection: entry into the targeted cells, break the defenses, survive and counteract acidification, which require to be understood at the molecular level. An important point to address is how the innate signalling affects Mtb intracellular trafficking.

1.6 Where do you think cellular microbiology is headed in the future? What do we need to be thinking about?

The field is moving towards ultra‐high resolution techniques. This would be fantastic to see the microbe and each of its components interacting with tissues and spreading within the host cells. It would be prodigious to be able to visualise in real time the action of a drug on its cellular targets. This would definitely contribute to a better understanding of the multiple actions of drugs.

1.7 What do you enjoy the most about your work? And what about the least?

I love the first step in discovery: finding new chemical Hits or new host susceptibility genes and reaching the proof of concept in vivo. Next, I enjoy deciphering the molecular and cellular mechanisms involving the new Lead or the cellular partners. I am grateful to my long‐time collaborators, in particular Laleh Majlessi and Edouard Yeramian, who have been helping me in my work.

On the less pleasant side, there is the administrative burden with regulations becoming more complex every day. I fear that we could reach a point when the experimental work will become overwhelmed by the time‐consuming regulatory requirements and administrative hurdles.

1.8 What has been the most surprising result you've had in your career so far?

When I was in Seoul, Laurent Marsollier came for a sabbatical year. While he had initially planned to do research on Mtb virulence, he went to explore the cellular mechanism of mycolactone, the main virulence factor of Mycobacterium ulcerans. He showed that mycolactone had an effect on intracellular calcium and potassium release within minutes. We went on to set‐up an RNAi screen to identify the pathways accounting for this effect and upon analysing the results we were literally puzzled by the list of putative genes we obtained. Most notably we were led to focus on the angiotensin 2 receptor 2 (AT2R) as a key player in the pathway. Subsequently it was a really thrilling time when we made the experiments, which demonstrated that indeed that AT2R was actually required for mycolactone signalling. Most importantly, this demonstration further allowed to elucidate the analgesia effect induced by mycolactone accounting for the painlessness of Buruli ulcer.

1.9 What advice would you give to other early career researchers starting out in their career given your experiences?

I would advise early career researchers to perform intensive experimental work and to care about serendipitous results. They should always try to use cutting‐edge techniques to move current paradigms forward. For instance, in the TB field, we tend to stick to old culture media, lengthy and cumbersome CFU plating (often asked by reviewers), younger ones should take risky roads and establish new niches as there are still so many unexplored traits of the TB bacillus.

Early career researchers should also discuss with the more experienced ones to avoid repeating the same mistakes. Results should be shared more openly and all results deserve to be published.

1.10 What has been your most rewarding experience so far? Why?

There have been many rewarding experiences. I can cite the Gordon Conference in Oxford in 2009 where I could show the success of the phenotypic screening for TB drug discovery. This has led to the initiation of many collaborations worldwide in Academia and in the Private and the development of many new preclinical candidates against TB.

中文翻译：

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细胞微生物专访

1 介绍

Priscille 在 Gustave Roussy 研究所攻读 HIV 整合酶药理学博士学位，然后在法国巴黎巴斯德研究所与 Stewart T. Cole 教授继续博士后研究。在成为 INSERM 研究员后，Priscille 于 2006 年移居首尔韩国巴斯德研究所，并在 Avenir 计划的支持下成立了她的团队。2011 年，Priscille 获得了 ERC-Consolidator 资助，并将她的团队迁往位于欧洲十字路口的法国里尔。她对致病性分枝杆菌如何与其宿主细胞相互作用以及小分子抑制剂如何阻止这些相互作用感兴趣。她对高内涵筛选充满热情，并乐于分享她的技术专长，以解决有关宿主-病原体相互作用的关键问题。

1.1 您的研究背景是什么——在您的学术生涯之初，您首先开始从事什么工作？

我在法国蒙彼利埃的化学工程学院学习生物学。我在硕士 2 期间进行了两次培训，第一次是关于通过 HIV TAT 肽递送寡核苷酸，第二次是关于男性决定因子 SRY 的双杂交方法。紧接着，我搬到了巴黎，在 Gustave Roussy 研究所攻读 HIV 整合酶抑制剂开发博士学位。1997年初，抗逆转录酶和抗蛋白酶抑制剂联合治疗HIV的多药疗法刚刚面世，前景广阔。因此，针对第三种 HIV 酶似乎是要走的路。最初的目标是在逆转录病毒基因组和寡核苷酸之间制造 DNA 三链体，以防止整合酶与其底物结合，我最终选择了适体！

1.2 你一直想在学术界工作吗？

其实没有。在获得博士学位后，我犹豫是否要加入一家生物技术公司。此外，我也曾在北美或法国的机会之间犹豫不决。然而，当我被邀请加入巴斯德研究所进行基于比较基因组学的结核病 (TB) 疫苗项目时，我非常激动，并毫不犹豫地加入了该实验室。斯图尔特科尔。事实上，我非常享受博士后的时光，经常对我的新生说博士后时期是研究生涯中最好的时期。巴斯德研究所对于许多研究主题来说确实是一个令人兴奋的地方，通过加入欧盟项目，我有机会与欧盟的不同实验室建立非常好的和富有成效的联系。那段时间，我申请了法国学术界的终身职位，并于 2005 年在 INSERM 取得了成功，所以我去了。

1.3 您早期的工作是如何将您带到现在的工作中的？

我一直很喜欢研究微生物并设计对抗它们的策略！我目前的研究主要是关于结核分枝杆菌 (Mtb)，我在博士后期间已经熟悉了。Roland Brosch 和 Alex Pym 教会了我所有的基础知识。然后，我在新成立的韩国巴斯德研究所获得了一个令人兴奋的研究机会，这是一个包括建筑设计过程在内的全新研究环境。在那里，我成功地开发了毒力Mtb 的表型分析适用于高通量筛选调节剂库的菌株。与蒂埃里·克里斯托夫 (Thierry Christophe) 一起采取的这些方法结果证明非常成功，例如发现了 Q203，目前是 IIB 期候选药物。回到法国后，我通过专注于破译新作用机制的研究项目来寻求新药。我们已将这些方法进一步扩展到其他细菌和微生物。值得注意的是，我们在锁定期间对 SARS-Cov2 感染的细胞进行了基于图像的表型筛选。

1.4 您一直对细胞微生物学感兴趣吗？

我一直对化学-微生物学感兴趣。我对组学中各种路径的探索感到非常兴奋。因此，在我的研究中实施比较和功能基因组学，当时是开创性的，导致发现了Mtb 中的7 型分泌系统. 我仍然清楚地记得当我看到第一只感染了 BCG 过表达 RD1（BCG-RD1）的 SCID 小鼠的表型时的惊讶！然后，我们在 3 年内筛选了 200,000 多种化合物，从而开发了 Q203。这无疑是我职业生涯中一个非常荣幸的时刻，因为我渴望为发现抗结核药物做出贡献。这是又一个激动人心的时刻，证明了新候选药物的体内功效！然后，我获得了 ERC 来研究基本问题，特别是针对Mtb 的宿主导向方法，这些方法在 2010 年被认为是先驱研究，但现在在微生物学领域非常流行。

1.5 你认为目前细胞微生物学中最有趣的问题是什么？

首先，结核病治疗依赖于服用几个月的药物组合。通常是四种药物。非常有效，但作用缓慢且容易受到抵抗。如何保持这种组合的有效性并避免出现耐药性和耐受性？我相信更好地了解施用这些抗生素后的宿主反应是关键的一部分。

其次，由于巨噬细胞生态位的重要性，Mtb与肺部和体内各种细胞的相互作用也受到了限制。局部肺上皮或感觉神经元信号也可能有助于巨噬细胞反应，这值得用新开发的工具（如类器官或肺芯片）进行探索。

第三，Mtb还使用多种策略来成功完成感染的各个步骤：进入目标细胞、打破防御、存活和抵消酸化，这些都需要在分子水平上加以理解。要解决的一个重要问题是先天信号如何影响Mtb细胞内运输。

1.6 您认为细胞微生物学未来的发展方向是什么？我们需要思考什么？

该领域正朝着超高分辨率技术发展。看到微生物及其每个成分与组织相互作用并在宿主细胞内扩散，这将是非常棒的。能够实时可视化药物对其细胞靶标的作用将是惊人的。这肯定有助于更好地了解药物的多种作用。

1.7 你最喜欢你的工作什么？最少的呢？

我喜欢发现的第一步：寻找新的化学命中或新的宿主易感基因，并在体内进行概念验证。接下来，我喜欢破译涉及新铅或细胞伙伴的分子和细胞机制。我感谢我的长期合作者，特别是 Laleh Majlessi 和 Edouard Yeramian，他们一直在我的工作中帮助我。

在不太令人愉快的一面，随着法规每天变得越来越复杂，行政负担越来越重。我担心我们可能会达到一个点，实验工作将被耗时的监管要求和行政障碍所淹没。

1.8 迄今为止，您职业生涯中最令人惊讶的结果是什么？

当我在首尔时，Laurent Marsollier 来休假一年。在他最初计划进行Mtb毒力研究的同时，他开始探索溃疡分枝杆菌的主要毒力因子分枝杆菌内酯的细胞机制. 他表明，分枝杆菌内酯在几分钟内对细胞内钙和钾的释放产生影响。我们继续设置了一个 RNAi 筛选来确定解释这种影响的途径，在分析结果时，我们对我们获得的推定基因列表感到困惑。最值得注意的是，我们被引导关注血管紧张素 2 受体 2 (AT2R) 作为该途径中的关键参与者。随后，当我们进行实验时，这是一个非常激动人心的时刻，这证明了实际上霉菌内酯信号需要 AT2R。最重要的是，该演示进一步阐明了由霉菌内酯引起的镇痛作用是布鲁里溃疡无痛的原因。

1.9 根据你的经历，你会给其他早期职业研究人员什么建议？

我会建议早期职业研究人员进行密集的实验工作并关心偶然的结果。他们应该始终尝试使用尖端技术来推动当前的范式向前发展。比如在TB领域，我们倾向于固守旧培养基，冗长繁琐的CFU电镀（审稿人经常问），年轻的应该走冒险的路，建立新的利基，因为TB还有很多未开发的特征杆菌。

早期职业研究人员还应该与更有经验的研究人员讨论，以避免重复同样的错误。结果应该更公开地分享，所有结果都应该公布。

1.10 到目前为止，您最有收获的经历是什么？为什么？

有很多有益的经历。我可以引用 2009 年在牛津举行的戈登会议，在那里我可以展示结核病药物发现的表型筛选的成功。这导致在全球学术界和私人领域发起了许多合作，并开发了许多新的抗结核临床前候选药物。