Epithelial plasticity in lung cancer evolution: insights from organoids

Sungji Moon; Dakyung Lee; Jaeyong Choi; Seyoon Lee; Jong-Il Kim

doi:10.51335/organoid.2026.6.e2

Organoid > Volume 6; 2026 > Article

Moon, Lee, Choi, Lee, and Kim: Epithelial plasticity in lung cancer evolution: insights from organoids

Review Article

Organoid 2026;6:e2.

Published online: February 25, 2026

DOI: https://doi.org/10.51335/organoid.2026.6.e2

Epithelial plasticity in lung cancer evolution: insights from organoids

Sungji Moon^1,^*

, Dakyung Lee^2,^3,^4,^*

, Jaeyong Choi⁵

, Seyoon Lee^2,^3,⁴

, Jong-Il Kim^2,^3,^4,⁶

¹Department of Social and Preventive Medicine, Sungkyunkwan University School of Medicine, Suwon, Korea

²Genomic Medicine Institute, Medical Research Center, Seoul National University, Seoul, Korea

³Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea

⁴Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, Korea

⁵Department of Biochemistry and Molecular Biology, Kangwon National University School of Medicine, Gangwon, Korea

⁶Cancer Research Institute, Seoul National University, Seoul, Korea

Correspondence to: Jong-Il Kim, MD, PhD Genomic Medicine Institute, Medical Research Center, Seoul National University, 103 Daehak-ro, Jongno-gu, Seoul 03080, Korea E-mail: jongil@snu.ac.kr

*These authors contributed equally to this work.

Received January 12, 2026 Accepted February 11, 2026

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Lung cancer remains the leading cause of cancer-related mortality and is often diagnosed at an advanced stage, limiting opportunities for early intervention. In vitro 3-dimensional lung organoid cultures recapitulate key features of human lung epithelial differentiation and are amenable to genetic manipulation. Epithelial plasticity during lung cancer evolution can be investigated by applying single-cell sequencing technologies to genetically engineered organoids, enabling the identification of mechanisms that suppress lung cancer initiation and progression. In this review, we consolidate recent findings and approaches in lung organoid–based cancer modeling and provide updated insights into lung cancer biology and future research directions.

Keywords: Lung neoplasms; Organoids

Introduction

Lung cancer is the leading cause of cancer-related death, accounting for approximately 1.8 million deaths worldwide in 2022 [1]. Because the disease often presents with limited and nonspecific symptoms in its early stages, timely diagnosis and intervention remain challenging [2]. Preventive and early-detection options are limited; smoking avoidance and low-dose chest computed tomography for high-risk individuals remain the primary strategies [3]. Molecular therapies that target specific genomic alterations, such as tyrosine kinase inhibitors, have improved survival among patients with lung cancer; however, drug resistance frequently develops over time [4]. The molecular mechanisms underlying lung cell plasticity during lung cancer development and progression remain incompletely understood [5]. Human lung cancer cell lines often lack features of normal cellular differentiation, making it difficult to model the earliest phases of oncogenesis. Accordingly, effective treatment and prevention strategies require models that accurately reflect disease pathogenesis. Genetically engineered mouse models (GEMMs) have provided valuable insights into lung cancer biology [6]; however, generating new GEMMs is resource-intensive, and these models can exhibit important physiological differences from humans.

In vitro 3-dimensional (3D) lung organoid cultures embedded in basement membrane extracts such as Matrigel simulate key aspects of human lung physiology and offer a promising alternative for studying lung cancer pathogenesis [7]. These organoids display differentiation potential within defined stem cell niches, closely recapitulate features of human lung epithelium, and can be readily manipulated genetically in vitro. Applying advanced next-generation sequencing approaches to these models can reveal previously uncharacterized mechanisms underlying lung cancer evolution. Accordingly, lung organoids have emerged as powerful tools in lung cancer research, serving both as validation systems for in vivo findings and as primary experimental platforms.

Several recent publications have reviewed general methodologies and applications of lung organoids [8]; cancer research using organoids derived from multiple tissues, including the lung [9,10]; murine lung organoid models for lung squamous cell carcinoma (LUSC) [11]; human lung cancer organoid systems [12,13]; studies of the cell of origin of lung cancers using in vivo models [14]; and broader work on lineage plasticity in cancer [15]. In this review, we focus specifically on lung organoid–based studies that use genetic engineering to model lung cancer, with particular emphasis on how gene alterations shape lung epithelial plasticity during disease initiation and progression. We summarize the genetic manipulation approaches used in these studies and interpret key findings in the context of recent in vivo discoveries on plasticity in normal lung differentiation and cancer development. Finally, we discuss current limitations and future perspectives for lung organoid research in cancer biology.

Lung stem cells: origin of differentiation and cancer

Ethics statement: This study was a literature review of previously published studies and was therefore exempt from institutional review board approval.

Specific lung cell types, lineages, and genetic alterations play critical roles in determining oncogenesis, tumor histology, and cellular transformation potential, which collectively influence prognosis in lung cancer [16–19]. Recent single-cell RNA sequencing studies using both in vivo and organoid models have clarified key trajectories of normal lung epithelial differentiation (Fig. 1A) [20–22]. Our study demonstrated that human airway organoids contain adult stem cells and their differentiated progeny, thereby reproducing the hierarchical organization of the human lung epithelium, including rare cell populations [20]. In the proximal lung, basal stem cells give rise to club cells through intermediate suprabasal cells [23]. Subsets of basal cells can also differentiate into rare epithelial cell types, including tuft cells, neuroendocrine cells, and ionocytes [20]. Club cells can further function as progenitors, differentiating into goblet cells or deuterosomal cells, which serve as precursors of multiciliated cells [24]. At the bronchioalveolar junction, bronchioalveolar stem cells can differentiate into either club cells or alveolar type 2 (AT2) cells [25]. In the distal lung, AT2 cells act as stem cells that differentiate into alveolar type 1 (AT1) cells, thereby maintaining the alveolar epithelium in the lung periphery [26].

Stem cells may be more prone to malignant transformation because their higher proliferative capacity increases opportunities to acquire mutations compared with more differentiated cells [27]. However, terminally differentiated cells can also serve as cells of origin for cancer if they undergo transdifferentiation or dedifferentiation after inflammation or injury—conditions that can trigger proliferation [28,29]. Because a narrow focus on stem cells as the cell of origin may overlook other potential sources of lung cancer, flexible genetic manipulation targeting multiple cell types within lung organoids provides an unbiased strategy to identify previously unrecognized mechanisms of cancer initiation, particularly when combined with cellular barcoding approaches [30,31]. Cancer initiation is determined not only by differentiation state but also by cell type–specific intrinsic gene programs and mutations [28], a concept referred to as molecular tropism [32]. Accordingly, organoid culture provides an ideal system for investigating cancer initiation because it enables modeling of lineage-specific organoids derived from distinct stem cells and their differentiated progeny.

Lung organoid types: in vitro stem cell culture

Lung organoids are 3D epithelial structures that self-organize in culture. These 3D systems can be expanded for more than six months without viral- or oncogene-mediated immortalization [33,34], allowing them to retain key features of normal lung epithelial biology. As a result, they provide an effective platform for monitoring the molecular and cellular transitions that occur during lung tumor development (Fig. 1B). Lung organoids can be generated from several sources, including adult lung stem cells (ASCs) [33–35], induced pluripotent stem cells (iPSCs) [36–38], and patient-derived organoids (PDOs) derived from samples obtained directly from lung cancer tissues.

1. Adult lung stem cells

Each lung stem cell type (basal cells, bronchioalveolar stem cells, or AT2 cells) can be isolated using fluorescence-activated cell sorting based on well-defined surface markers. Alternatively, whole lung tissue can be enzymatically digested and the resulting cell mixture cultured in stem cell–supportive media, enriching stem cells without the need for sorting. Lung organoids derived from GEMMs are commonly used to study lung cancer because they often carry engineered genetic alterations introduced at the embryonic stage (Table 1) [39–57]. In addition, ASCs isolated from normal human or mouse lungs can be modified in vitro using genome engineering tools to create experimental lung cancer models.

1) Induced pluripotent stem cells

Human iPSCs can first be directed into lung progenitor states and then guided to form airway organoids [16], alveolar organoids [37], or mixed-lineage structures containing both compartments [18]. Beyond epithelial differentiation, iPSCs can also be directed toward mesoderm-derived cell types (e.g., endothelial cells, fibroblasts, and immune lineages), enabling co-culture systems that more closely approximate the native lung microenvironment. iPSC-derived alveolar organoids have been widely applied to study lung adenocarcinoma (LUAD), whereas iPSC-based airway organoids are used far less frequently in cancer research. As in mouse models in which genome editing is performed at the embryonic stage, genetic modifications can be introduced into iPSCs before differentiation to create engineered cancer models.

2) Patient-derived organoids

PDOs can be expanded in 3D culture while retaining each patient’s distinct genetic and non-genetic alterations, providing a powerful platform for dissecting individualized cancer biology and therapeutic vulnerabilities [58]. These organoids can also be genetically engineered to probe mechanisms of phenotypic change, including shifts in histologic identity. Multiple studies have leveraged lung cancer PDO models to examine such phenotype transitions [44,56,57].

Genetic engineering methods

To model tumor initiation and progression, lung organoids can be engineered to introduce loss-of-function alterations in tumor suppressor genes or gain-of-function alterations in oncogenes. In addition, several genome manipulation tools enable spatiotemporal control of target gene expression. Published protocols describe CRISPR-based genome editing [59] and electroporation-based transfection in organoid cultures [60]; these methods can be adapted to specific experimental needs.

1. Loss-of-function mutation

Cas9 produces double-strand breaks directed by single-guide RNAs (sgRNAs), and subsequent non-homologous end joining generates insertions and deletions that can disrupt tumor suppressor genes, resulting in gene knockout (KO) [61]. sgRNAs are typically designed to target regions near the start of the coding sequence or exons essential for gene function. Gene knock-in (KI) can be achieved through homology-directed repair (HDR) using a donor DNA template, enabling the introduction of specific point mutations that inactivate tumor suppressor genes [36]. Compared with HDR-based KI, base editing and prime editing can provide more efficient approaches for introducing precise point mutations at the desired genomic locus [59]. For example, Ebisudani et al. [44] introduced Cas9 and sgRNAs targeting TP53 or CDKN2A into alveolar organoids using nonviral delivery vectors.

2. Gain-of-function mutation

KI approaches can also be used to introduce gain-of-function mutations at endogenous proto-oncogene loci. Ectopic gene cassettes may be integrated into so-called “safe harbor” loci—such as Rosa26, H11, and Col1a1 in the mouse genome, or AAVS1 and CLYBL in the human genome—which are considered permissive sites that do not disrupt endogenous gene transcription. Oncogene overexpression can be achieved through KI strategies or by introducing complementary DNA constructs into organoids using lentiviral vectors or nonviral gene delivery platforms, including the PiggyBac transposon system. In addition, oncogenic fusion events can be generated experimentally. For example, Naranjo et al. [46] induced the EML4–ALK fusion in murine alveolar organoids by delivering sgRNAs that create targeted chromosomal breaks in each partner gene, thereby promoting translocation [32]. Similar strategies have been applied to generate other oncogenic rearrangements in lung models. Choi and Meyerson [62], for instance, produced de novo CD74–ROS1 and KIF5B–RET fusions in vivo using sgRNAs designed to induce specific chromosomal translocations.

3. Gene expression control

Gene expression in organoid systems can also be regulated using Cre–LoxP–based strategies. Flanking a gene with LoxP sites enables conditional knockout, whereas conditional expression can be achieved by inserting a LoxP-stop-LoxP (LSL) cassette upstream of the transcriptional start site. Alleles such as Trp53^f/f and LSL-Kras^G12D/+ are commonly used in LUAD models. Cre recombinase recognizes LoxP sites and mediates site-specific recombination. For example, Hai et al. [53] introduced sgRNAs targeting Trp53, Pten, and Cdkn2a together with Cre recombinase into mouse airway organoids harboring LSL-Sox2 in Rosa26 and LSL-Cas9 in H11 using lentiviral delivery.

Cre activity can itself be externally controlled. Adenovirus expressing Cre (Ad-Cre) induces recombination at LoxP sites and thereby modulates gene expression. Temporal control can be achieved using tamoxifen-inducible Cre variants, such as Cre^ERT or Cre^ERT2. In these systems, the modified estrogen receptor retains Cre recombinase in the cytoplasm until tamoxifen administration triggers nuclear translocation and recombination. Another inducible strategy relies on tetracycline-responsive promoter elements. The reverse tetracycline transactivator (rtTA) activates gene expression from tetracycline-responsive promoters in the presence of doxycycline. For example, Dost et al. [39] inserted TetO-KRAS^G12D and rtTA into the AAVS1 locus of iPSCs to regulate KRAS expression in alveolar organoids. Cell type–specific promoters further refine gene control. Promoters such as SFTPC^CreER for AT2 cells [42], KRT5^CreER for basal cells [50], and ASCL1^CreER for neuroendocrine lineages enable lineage-restricted induction of genetic alterations.

4. In vitro mutated organoid selection

When organoids are genetically modified in vitro, edited populations can be enriched based on the introduced molecular alterations. Fluorescent reporters and antibiotic resistance genes are commonly used to isolate organoids that have successfully acquired the intended mutations. Single-clone–derived mutant organoids can be selected by manual picking [59], whereas analysis of multiple clones enables assessment of polyclonal effects [53]. In some cases, the genetic alteration itself confers a selective growth advantage. Organoids with oncogenic pathway activation may proliferate under conditions in which normal organoids cannot, such as media lacking essential niche factors [61]. Ebisudani et al. [44] demonstrated that withdrawal of specific growth factors suppressed expansion of wild-type alveolar organoids, whereas TP53- or CDKN2A-deficient organoids and those expressing EGFR^L858R or KRAS^G12D continued to proliferate robustly.

Mutant organoids can also be enriched through small-molecule–based functional selection. For example, TP53-null organoids exhibit resistance to nutlin-3a, an MDM2 inhibitor that selectively eliminates p53-proficient cells [61,63,64]. Similarly, RB1-deficient organoids can be maintained in the presence of palbociclib, a CDK4/6 inhibitor [64]. Additional mutation-specific selection strategies have been described by Celotti et al. [59].

Strengths of organoids compared with in vivo models

Lung organoids provide a versatile platform for precise genetic manipulation and often represent a more practical alternative to generating new GEMMs. They permit combinations of gene alterations that may be technically challenging to introduce simultaneously in vivo. When targeted gene control is difficult to achieve in GEMMs, organoid systems offer a valuable substitute because epithelial cells can be directly modified in vitro. For example, adenoviral delivery of genes to basal cells in the upper airway is limited by mucociliary barriers [50], whereas basal cell–derived airway organoids can be efficiently transduced in culture.

Shared progenitor lineages across tissues can also complicate the development of lung-specific GEMMs. A model driven by KRT5-specific oncogenic activation failed to generate lung tumors because KRT5-positive basal cells in the skin developed tumors first [50]. Jeong et al. [50] addressed this limitation by isolating tracheal epithelial cells from the KRT5-specific GEMM and manipulating them in vitro. A similar challenge arose in an Ascl1-driven model in which mice developed thyroid tumors; Gardner et al. [47] therefore used airway organoids derived from the GEMM to investigate lung-specific tumorigenesis.

Certain gene-editing strategies are further constrained by the genomic architecture of the mouse. For example, achieving AT2-specific deletion of Rb1 would require homozygous floxing of Rb1 combined with Sftpc-driven Cre expression. However, the close proximity of Rb1 and Sftpc on chromosome 14 (approximately 2.6 Mb apart) increases the risk of unintended recombination events [47]. A practical alternative is to use alveolar organoids derived from GEMMs carrying LSL-Cas9 and ^Sftpc-CreERT2, followed by plasmid-based delivery of sgRNAs targeting Rb1.

Organoid-based systems are particularly valuable when mouse models fail to recapitulate critical aspects of human disease. Qu et al. [65] addressed the inability of murine small cell lung cancer (SCLC) models to metastasize to the brain by combining murine SCLC organoids with human iPSC-derived cortical organoids to investigate mechanisms of brain metastasis. This strategy overcame a key limitation of mouse SCLC models, which rarely develop brain metastases despite the high clinical incidence in patients [65]. Similarly, Liu et al. [66] used PDO models in vivo to study metastatic lung cancer biology, addressing the limitation that LUAD patient-derived xenograft models infrequently produce metastatic lesions [67].

Characterization of phenotype transition in vitro and in silico

1. In vitro capture

Once genetic alterations are introduced, lung organoids can be longitudinally monitored to assess phenotypic changes. Bright-field microscopy enables quantitative evaluation of morphological features, including organoid size, circularity, wall thickness, and transitions from smooth, cystic structures to irregular, solid morphologies—features that often accompany oncogenic transformation [14]. Proliferative capacity can be assessed using organoid formation assays or by measuring proliferation markers such as Ki-67 through immunohistochemistry or quantitative reverse transcription polymerase chain reaction. Cellular viability can be evaluated using metabolic assays, including water-soluble tetrazolium salt-1 (WST-1) or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, which measure mitochondrial activity.

The definitive assessment of malignant transformation involves transplantation of genetically modified organoids into mouse models. Orthotopic implantation into the murine lung provides an in vivo microenvironment that more closely resembles human lung cancer development [54]. Histologic evaluation of organoids or tissues derived from in vivo transplants can be performed using hematoxylin and eosin staining in combination with lung cancer subtype–specific markers (e.g., TTF1 or NKX2-1 for LUAD; ΔNp63 or KRT5 for LUSC; ASCL1, NEUROD1, POU2F3, or YAP1 for SCLC).

2. In silico capture by single-cell sequencing

Single-cell RNA sequencing enables transcriptomic profiling at single-cell resolution, capturing cellular heterogeneity that cannot be resolved by bulk sequencing. This approach can identify intermediate cellular states that emerge during lung cancer progression, as these transitional populations display distinct gene expression signatures (Fig. 1B). Transformed cancer cells typically exhibit increased cell-cycle activity, elevated expression of lineage-associated oncogenic markers, and more pronounced copy number variation (CNV) relative to normal epithelial cells. CNVs, a hallmark of cancer, can be inferred from single-cell transcriptomic data using computational tools such as inferCNV [68], CopyKAT [69], or Numbat [70]. Numbat integrates haplotype information with RNA expression data, enabling more accurate CNV inference than approaches that rely solely on expression patterns [70].

Trajectory-based analyses, including CellRank [71], Monocle [72,73], and partition-based graph abstraction [74], can delineate transitional states during malignant evolution and help infer potential cells of origin. These approaches often incorporate RNA velocity, which estimates transcriptional directionality by comparing unspliced and spliced RNA transcripts using tools such as Velocyto [75] or scVelo [76]. Gene regulatory networks underlying these transitions can be reconstructed using frameworks such as SCENIC [77] or TENET [78], which infer transcription factor activity and downstream regulatory interactions.

These transcriptional dynamics are closely linked to epigenomic remodeling. Chromatin accessibility varies across cell types and can be assessed using single-cell assay for transposase-accessible chromatin sequencing (ATAC-seq) [79] or through multimodal approaches that simultaneously profile RNA expression and chromatin accessibility. Time-resolved single-cell RNA or ATAC sequencing further enables investigation of dynamic regulatory processes along developmental or oncogenic trajectories. In silico gene perturbation analyses based on single-cell RNA sequencing data from lung organoids—using tools such as CellOracle [80], scTenifoldKnk [81], or SCENIC+ [82]—can provide insights into the effects of specific gene perturbations on normal lung differentiation.

Transformation from normal organoids

Lung cancer is histologically classified into SCLC and non-SCLC (NSCLC), the latter comprising LUAD, LUSC, and large-cell carcinoma. In practice, mixed histologic subtypes such as adenosquamous carcinoma and large-cell neuroendocrine carcinoma are also observed. These mixed phenotypes may arise from shared lineage populations, a hypothesis that can be effectively examined using lung organoid models. Distinct genomic mutation patterns characterize each histologic subtype, although TP53 mutations are common across lung cancer types [83,84]. Frequently mutated genes in human lung cancer therefore represent key targets for investigating their functional roles in tumor initiation and progression. In NSCLC, oncogenic mutations frequently contribute to therapeutic resistance, ultimately leading to treatment failure [5]. One mechanism underlying resistance in LUAD is histologic transformation, in which tumors adopt features of SCLC or LUSC; such transitions can also be modeled and studied using organoid systems [56,57].

1. LUAD

1) Target genes and cell of origin

LUAD is frequently driven by single-gene alterations in EGFR, KRAS, BRAF, ALK, ROS1, and RET, all of which regulate intracellular signaling pathways associated with cellular proliferation [49]. In vivo studies indicate that LUAD predominantly arises from AT2 cells [16], and most LUAD-focused organoid investigations therefore use alveolar organoid models. These studies have primarily examined KRAS^G12D and EGFR^L858R mutations, often in combination with TP53 or CDKN2A loss. Additional alterations—including EML4–ALK fusions, BRAF^V600E [46], and HER2 overexpression [43]—have also been investigated. The Gprc5a^−/− mouse model spontaneously develops KRAS-mutant LUAD following carcinogen exposure [85]. Using organoids derived from Gprc5a^−/− mouse lungs at transitional stages, Han et al. [42] investigated early events in LUAD tumorigenesis.

2) LUAD from AT2 cells

Transformation of AT2 cells by mutant KRAS induces intermediate transitional cell states characterized by heightened plasticity, as demonstrated by single-cell RNA and ATAC sequencing analyses [86]. AT2 cells expressing KRAS^G12D lose their lineage identity [39] and acquire hybrid phenotypes marked by co-expression of AT1 and AT2 markers [40], along with reactivation of fetal lung developmental genes such as SOX9 [39] and activation of injury- or plasticity-associated gene programs [41]. The KRAS–ITGA3–SRC signaling axis has been identified as a critical mediator of this process and can be pharmacologically inhibited using SRC inhibitors [41]. Recently, Li et al. [87] performed multi-omics profiling (single-cell RNA and ATAC sequencing) of AT2 cells harboring mutant Kras and Trp53 and identified two subpopulations, one of which was more strongly associated with LUAD progression. In lung organoids derived from Gprc5a^−/− mice, KRT8-positive intermediate cells emerged prior to overt LUAD formation and exhibited reduced differentiation capacity with increased cellular plasticity [42]. Similarly, HER2 overexpression in iPSC-derived AT2-like cells induces atypical adenomatous hyperplasia, a recognized precursor lesion of LUAD [43]. The emergence of highly plastic, developmentally reminiscent cell states is a recurring feature of cancer [88,89]. Activation of lineage stage–specific intracellular signaling pathways may therefore reactivate fetal-like developmental programs. Future studies should examine whether other oncogenic alterations, including EGFR, BRAF, or ALK mutations, induce comparable effects in AT2 cells.

3) Lepidic adenocarcinoma from AT1 cells

Using an in vivo model, Juul et al. [90] demonstrated that activation of KRAS^G12D in AT1 cells—terminally differentiated descendants of AT2 cells—following lung injury results in the development of lepidic adenocarcinoma. In this context, LUAD proliferates along the alveolar surface through transdifferentiation of AT1 cells into AT2-like states. These findings indicate that fully differentiated cells can also serve as cells of origin for cancer. However, it remains unclear whether other oncogenic mutations can similarly reprogram differentiated cell types under injury conditions.

4) Wnt dependency in LUAD

Ebisudani et al. [44] reported that LUAD PDOs can be stratified into Wnt-dependent and Wnt-independent subtypes. Activation of EGFR^L858R or KRAS^G12D in human alveolar cells with TP53 and CDKN2A knockout induced Wnt independence, and the alveolar lineage transcription factor NKX2-1 was identified as a key regulator of this process [44]. Modulation of NKX2-1 expression in LUAD influenced tumor cell behavior and maintenance [44].

2. LUSC

1) Target genes and cell of origin

LUSC is typically characterized by multiple genomic alterations, most prominently amplifications on chromosome 3q [49]. Airway organoids derived from basal cells, which are considered the principal cell of origin for LUSC [17], are commonly used to model LUSC tumorigenesis. Among the genes investigated, SOX2—a transcription factor essential for cell fate determination during embryogenesis and a central regulator of squamous cell carcinoma—has been most extensively studied in airway organoids [91]. In addition to SOX2 overexpression, Liu et al. [52] introduced several 3q-amplified genes, including PRKCI and ECT2, whereas Hai et al. [53] incorporated additional mutations in Pten, Cdkn2a, and Trp53, all of which are frequently altered in LUSC. The roles of Bcl11a [51], Keap1 [50], and Kmt2d [54] in LUSC development have also been explored using organoid-based models.

2) LUSC from airway organoids

In airway-derived organoids, SOX2 overexpression suppresses mucociliary differentiation [92], whereas BCL11A expression—under SOX2 regulation—induces a squamous-like phenotype [51], representing a precursor state of LUSC. However, SOX2 expression alone is insufficient to fully transform basal cells into LUSC; instead, it results in precancerous lesions or squamous metaplasia [92,93]. Complete transformation to LUSC requires multiple cooperating genetic alterations, which can be effectively modeled in airway organoids. Liu et al. [52] demonstrated cooperative interactions among SOX2, PRKCI, and ECT2—all located on chromosome 3q26—in murine airway organoids. The protein product of PRKCI (PKCι) directly phosphorylates SOX2 and ECT2, thereby promoting squamous transformation. Additional genes co-amplified within the 3q region, including TP63 (3q28) and PIK3CA (3q26), have also been implicated in tumorigenesis. TP63 functions as a basal stem cell fate regulator, whereas PIK3CA acts as a proto-oncogene driving proliferation, migration, and invasion.

The high frequency of 3q amplification across squamous carcinomas of the lung, head and neck, esophagus, and cervix remains incompletely understood. Future organoid studies could systematically examine functional combinations of 3q-amplified genes. For example, AGTR1 (3q24), which encodes a G protein–coupled receptor targeted by angiotensin II, is co-amplified within this region. Epidemiological evidence suggests that angiotensin II receptor blockers may be associated with reduced lung cancer incidence [94]. Although AGTR1 has been linked to cancer cell proliferation, its role in LUSC tumorigenesis has not yet been experimentally examined. Additionally, combined Pten, Trp53, and Cdkn2a mutations with Sox2 overexpression in murine airway organoids can induce LUSC [53]. These findings suggest that although basal cells possess stem-like properties that may predispose them to oncogenic transformation, intrinsic tumor-suppressive mechanisms may delay malignant conversion. This may partially explain the 10–30-year latency period of lung cancer development observed among chronic smokers [95]. Pan et al. [54] further demonstrated that deletion of Kmt2d and Trp53 in murine airway organoids triggered LUSC through chromatin-mediated downregulation of protein tyrosine phosphatases, resulting in activation of EGFR and ERBB2 signaling.

Airway organoids contain multiple epithelial cell types, including club cells, which have also been shown to initiate LUSC in vivo [14]. Cell type–specific genomic perturbation therefore provides a powerful strategy to dissect differential susceptibilities among airway epithelial populations. For example, in basal cell–derived organoids, Keap1/Trp53 mutations induce squamous transformation specifically in basal cells when driven by Krt5-CreERT2, but not in differentiated secretory or mucociliary cells [50]. These observations underscore that oncogenic potential is closely linked to cellular identity and differentiation state. An important unresolved question is whether basal cells can revert to a more primitive, undifferentiated state—analogous to the transition of AT2 cells through intermediate states during regeneration and oncogenic transformation. Elucidating such dedifferentiation trajectories in basal lineages may further clarify the cellular origins and plasticity underlying LUSC initiation.

3) LUSC from LUAD

Tong et al. [56] demonstrated that LUAD organoids generated by Kras^G12D activation combined with Lkb1 loss can give rise to squamous carcinoma, and that resistance to Kras inhibition is mediated through the ELF5–ΔNp63 axis. Because oncogenesis was induced using Ad-Cre under a ubiquitous promoter, the precise cell of origin could not be definitively established. However, the findings suggest that club cells may contribute, as they have been shown to initiate both LUAD and LUSC in vivo [16,96].

3. SCLC

1) Target genes and cell of origin

SCLC is primarily driven by RB1 loss in combination with TP53 mutation [50]. In vivo studies have identified pulmonary neuroendocrine cells as the principal cell of origin for SCLC [97]. Transcriptionally, SCLC can be classified into four subtypes: SCLC-A, SCLC-N, SCLC-P, and SCLC-Y. The neuroendocrine subtypes SCLC-A and SCLC-N are defined by high expression of ASCL1 and NEUROD1, respectively [98]. In contrast, the non-neuroendocrine subtypes SCLC-Y and SCLC-P are characterized by elevated expression of YAP1 and POU2F3 [98]. MYC serves as a major driver of SCLC progression and cooperates with TP53 and RB1 loss to promote tumor development [99].

2) SCLC from airway organoids

Airway organoids contain neuroendocrine cells that arise from basal progenitors—rare neurosensory epithelial cells considered potential cells of origin for SCLC [18,100]. Ouadah et al. [101] demonstrated that specific neuroendocrine cell clusters acquire stem-like self-renewal capacity following injury, suggesting that they may represent authentic cells of origin for SCLC. Park et al. [55] modeled SCLC by inactivating RB1 and TP53 while overexpressing MYC, BCL2, and myristoylated AKT in human bronchial epithelial organoids. However, that study did not definitively establish whether transformation originated from neuroendocrine cells or from another epithelial subset. Gardner et al. [47] addressed this question by selectively targeting neuroendocrine cells in airway organoids using Ascl1-CreERT2 to drive MYC expression in combination with Trp53 and Rb1 loss, successfully inducing SCLC formation. They further demonstrated that basal cells can be reprogrammed into neuroendocrine-lineage cancers through Trp53/Rb1 knockout or Myc overexpression in vivo, whereas other lung epithelial cell types were resistant to Myc-driven transformation [47]. Although neuroendocrine cells originate from basal progenitors, it remains unclear whether SCLC arises from fully differentiated neuroendocrine cells that subsequently acquire mutations or from mutation-bearing basal cells that aberrantly differentiate toward a neuroendocrine lineage.

3) Origin of SCLC-P

Although MYC is a key driver of SCLC-A and promotes progression toward SCLC-N and SCLC-Y from neuroendocrine cells [99], the developmental origin of SCLC-P remains incompletely defined. It has been proposed that SCLC-P may arise from tuft cells, a rare lung epithelial population characterized by POU2F3 expression. Consistent with this hypothesis, Ireland et al. [102] demonstrated that SCLC-P can originate from basal cells following Trp53/Rb1 knockout combined with MYC overexpression in basal cell–derived murine airway organoids. Given that tuft cells, like neuroendocrine cells and ionocytes, arise from basal progenitors, basal cells harboring TP53 or RB1 mutations may generate SCLC-A upon differentiation into neuroendocrine cells and SCLC-P upon differentiation into tuft cells. Such lineage plasticity could contribute to the marked intratumoral heterogeneity observed in SCLC.

4) Novel drivers of SCLC

Chan et al. [103] identified PLCG2 as a gene broadly expressed across SCLC cell clusters based on single-cell RNA sequencing, implicating a potential role in SCLC metastasis. Functional interrogation of PLCG2 in normal airway organoids could clarify whether it also contributes to SCLC initiation. Additionally, Lee et al. [104] reported Tsc1 as a tumor suppressor involved in SCLC pathogenesis. Loss or dysregulation of Tsc1 could therefore be modeled in airway organoids to further elucidate its role in tumor initiation and progression.

5) SCLC from LUAD and LUSC

Gardner et al. [47] demonstrated in vivo that LUAD originating from AT2 cells driven by EGFR^L858R, followed by Trp53/Rb1 loss and Myc overexpression, can transition into a neuroendocrine carcinoma phenotype [22]. This transformation appears to involve an intermediate undifferentiated, basal-like state and circumvents the intrinsic resistance of normal AT2 cells to direct SCLC conversion. In contrast, histologic transformation from LUSC to SCLC has not yet been explored using lung organoid systems. Given that SCLC is highly aggressive, frequently metastatic, and often emerges as a transformed state from other lung cancer subtypes, it represents one of the most undifferentiated and plastic forms of lung cancer.

6) Hypothetical alternative sources of SCLC

High-grade ovarian cancer can originate from pre-ciliated tubal epithelial cells harboring Trp53 and Rb1 mutations, whereas progenitor cells carrying the same mutations undergo apoptosis [105]. By analogy, in the lung, deuterosomal cells—precursors of multiciliated cells—may represent a potential alternative cell of origin for SCLC driven by TP53 and RB1 inactivation. CDK4/6 phosphorylates RB1, and its role in multiciliogenesis has been examined [106,107]; however, the direct impact of RB1 deletion on multiciliated cell differentiation and its possible link to SCLC remain undefined. Primary cilia have been detected in SCLC specimens, whereas they are rarely observed in NSCLC [108]. Cochrane et al. [109] demonstrated that loss-of-function mutations in Trp53 and Rb1 suppress autophagy, thereby increasing primary cilia formation. Jain et al. [110] further showed that primary ciliated airway epithelial cells can give rise to motile ciliated cells. Notch signaling—one of the most frequently altered pathways in SCLC—plays a central role in cell fate determination: Notch inhibition promotes multiciliated differentiation from club cells and neuroendocrine differentiation from basal cells. Collectively, these observations suggest a potential mechanistic intersection between multiciliogenesis and SCLC evolution in the context of TP53 and RB1 loss, warranting further investigation into how ciliary differentiation programs may contribute to SCLC initiation.

Distinct susceptibility to transformation by cell type and mutation

1. Distinct susceptibility by cell type

KRAS^G12D can transform AT2 cells in the alveoli or club cells in the airway but fails to transform other bronchiolar epithelial cell types in vivo [16]. Similarly, mutant EGFR drives transformation in AT2 cells but not in basal or club cells, as demonstrated in organoid models [48]. SOX2, a key basal cell marker, suppresses KRAS-induced transformation by inhibiting Notch signaling [40], suggesting a context-dependent protective effect in LUAD [40]. Conversely, aberrant SOX2 expression promotes basal cell transformation toward LUSC while inhibiting progression of AT2 cells into LUAD [40]. Notably, SOX2 exhibits context-dependent roles across cancers—functioning as a tumor suppressor in gastric cancer while acting as a proto-oncogene in esophageal and lung cancers [111]. Even within lung cancer, SOX2’s function depends on cellular lineage: it promotes oncogenesis in basal cells but suppresses transformation in AT2 cells. Moreover, identical genetic alterations can give rise to distinct histologic subtypes depending on the cell of origin. For example, combined Keap1 and Trp53 deletion induces LUAD when introduced into peripheral alveolar cells but results in LUSC when induced in proximal airway basal cells [50].

2. Distinct susceptibility by mutation

Club cells function not only as a cell of origin for LUSC (along with basal cells) but also for LUAD within the airway, particularly when harboring oncogenic mutations such as KRAS^G12D or EML4–ALK [112,113]. In EML4–ALK–driven LUAD, club cells act as the predominant cells of origin and undergo transformation toward a squamous-like phenotype through activation of JAK–STAT signaling [57]. This susceptibility may reflect their developmental proximity to AT2 cells, as both lineages derive from bronchioalveolar stem cells, and club cells can differentiate into AT2 cells in response to injury [114]. Compared with basal cells, club cells likely express lower levels of SOX2, potentially rendering them more susceptible to LUAD transformation following MAPK pathway activation [115]. Additionally, Trp53 and Pten mutations induce LUAD formation in murine airway organoids [54], further supporting the hypothesis that club cells may serve as a plausible cell of origin for LUAD in this context. Chen et al. [48] demonstrated that bronchioalveolar stem cells transformed by EGFR mutations (L858R or T790M) can generate tumors with mixed adenomatous and squamous histology, exhibiting differential sensitivity to tyrosine kinase inhibitors depending on the original cell lineage. Collectively, these findings underscore that the cell of origin critically influences both histologic trajectory and therapeutic response in lung cancer.

3. Role of inflammation in susceptibility to tumorigenesis

Exogenous inflammatory stimuli can promote tumorigenesis by amplifying the oncogenic effects of pre-existing genetic alterations [116]. Hill et al. [45] demonstrated that exposure to particulate matter 2.5 (PM_2.5) enhances the oncogenic activity of EGFR^L858R in vivo and in lung organoid models. Inflammation may increase cellular susceptibility to malignant transformation, particularly given that mutated yet histologically normal epithelial cells are frequently detected in individuals without overt cancer [117,118]. Whether driven by intrinsic processes or environmental exposures, inflammation plays a pivotal role in lung tumorigenesis [28]. Because fusion oncogene–driven transformations, such as EML4–ALK in LUAD, often exhibit prolonged latency before clinical manifestation [119], environmental factors may facilitate oncogenesis by epigenetically remodeling chromatin architecture in ways that favor transformation. Lung organoids therefore provide a powerful experimental system for dissecting the biological effects of environmental carcinogens, including PM_2.5 and cigarette smoke, on lung epithelial cells. These models enable real-time observation of cellular and lineage-level responses in vitro. Investigating whether injury-induced transdifferentiation or dedifferentiation occurs during lung tumorigenesis—and whether such reprogrammed cells acquire stem-like properties—represents a promising direction for future organoid-based research on cancer evolution.

Limitations

Although lung organoid cultures grown within defined stem cell niches provide powerful systems for investigating human pulmonary epithelial biology and the evolutionary dynamics of lung cancer, several inherent limitations must be acknowledged [120].

1. Lung microenvironment

In vitro lung organoid models cannot fully recapitulate the structural and functional complexity of native lung tissue or its tumor microenvironment. Critical stromal and immune components—such as endothelial cells, fibroblasts, and diverse immune cell populations—play essential roles in lung physiology and cancer progression by interacting with epithelial cells and supporting the expansion of mutated clones. These interactions are largely absent in conventional organoid systems [121]. For example, in vivo, mutant epithelial cells may be eliminated through immune surveillance; in contrast, organoids lacking immune components do not replicate this clearance mechanism, potentially shortening the latency period for transformation. Conversely, immune and stromal cells in the native lung can also promote tumor progression by secreting growth factors and cytokines or by remodeling the extracellular matrix (ECM). Accordingly, while organoids offer valuable mechanistic insight into epithelial biology and oncogenic transformation, they do not fully model the cellular diversity, spatial organization, or dynamic intercellular interactions that shape lung cancer evolution in vivo.

2. Reproducibility

Matrigel, commonly used as the ECM for lung organoid culture, is derived from murine sarcoma and may not accurately reflect the biochemical or mechanical properties of human ECM. In addition, genomic instability that may arise during prolonged culture has not been systematically characterized in lung organoids derived from either iPSCs or ASCs, potentially affecting reproducibility. Adult stem cell–derived lung organoids are often established from histologically normal tissue adjacent to tumors in patients with lung cancer and may therefore harbor somatic mutations. Conversely, PDOs generated directly from lung cancer tissue frequently face overgrowth by non-malignant epithelial organoids, necessitating additional purification steps to selectively enrich cancer-derived populations [122].

Further applications

1. Co-culture

Co-culture of lung organoids with mesoderm-derived cell types—such as fibroblasts, endothelial cells, and diverse immune populations—can generate more physiologically relevant models of the tumor microenvironment during lung cancer progression. Organoid-on-a-chip platforms further extend this approach by recreating systemic, multi-tissue interactions within microfluidic devices [123]. These systems may complement or partially replace certain in vivo models by enabling dynamic tracking of metastatic dissemination, including migration of cancer cells from lung organoids to distant organ compartments within the same platform.

Another emerging strategy involves the generation of “assembloids,” in which two or more distinct organoid types are integrated. This approach is applicable to lung organoids; for example, Qu et al. [65] combined SCLC organoids with human iPSC-derived cortical organoids to investigate mechanisms of brain metastasis.

2. Genetic engineering technology

The adaptability of lung organoids to diverse genetic engineering strategies offers opportunities to identify previously unrecognized mechanisms of lung cancer initiation and progression. Pooled sgRNA library screens can be performed in organoid systems to uncover novel driver mutations. Many genomic alterations associated with lung cancer—including variants in noncoding regions—are difficult to interrogate functionally in GEMMs. In contrast, organoid platforms are well suited for high-throughput functional assays, such as massively parallel reporter assays, enabling systematic evaluation of these variants.

Population-specific mutation patterns further underscore the need for human-relevant models. For example, EGFR mutations in NSCLC occur more frequently in East Asian populations (49.1%) than in European populations (12.8%) [124]. Studying alveolar organoids derived from individuals of diverse ancestries may help clarify biological or genetic factors underlying these disparities—an aspect that cannot be adequately modeled in mice. Advanced genome engineering tools—including CRISPR-Cas9, base editing, prime editing, and transcriptional regulation using catalytically inactive Cas9 (dCas9)—provide flexible approaches for modifying genes of interest in organoids. Novel genomic insights derived from single-cell RNA or ATAC sequencing of lung cancer can be functionally evaluated in human lung organoids with relative ease [79,125,126].

3. Sequencing technology

Elucidating the cellular origins of lung cancer and the differentiation disruptions that accompany tumor evolution requires lineage-tracing strategies. These can be addressed not only through computational inference but also through experimental approaches such as molecular barcoding [118,119]. Because lung organoids grow in 3D structures, they are amenable to spatial transcriptomics and spatial proteomics, enabling reconstruction of spatiotemporal lineage dynamics at single-cell resolution and characterization of tumor–microenvironment interactions over time. Single-cell genome sequencing approaches, including Strand-seq [127], allow investigation of clonal evolution and can detect mosaic structural variants that arise during transitional stages of tumorigenesis [128]. Additional computational tools for detecting somatic mutations at single-cell resolution—such as Monopogen [129] and Scomatic [130]—can further delineate subclonal architecture.

Beyond chromatin accessibility profiling using single-cell ATAC-seq, additional epigenomic modalities—including Hi-C for 3D genome architecture and DNA methylation profiling—can be applied to organoid systems. Integrative approaches that combine these layers at single-cell resolution, such as single-nucleus methyl-3C sequencing [131], enable comprehensive mapping of epigenomic remodeling during malignant transformation. Single-cell proteomics provides complementary information to transcriptomic analyses, as mRNA abundance does not always correlate with protein expression and many regulatory processes operate through post-translational modifications that are not captured by RNA-based methods [132]. Finally, long-read DNA and RNA sequencing technologies facilitate the detection of structural variants and alternative RNA isoforms, offering additional insight into patient-specific trajectories of lung cancer evolution.

4. Non-invasive early detection

Detection of extracellular biomarkers—such as metabolites, exosomes, circulating cell-free DNA, or microRNAs—through liquid biopsy approaches is typically more effective in patients with advanced tumor burden. Investigating these biomarkers in culture media from lung organoids during early stages of tumorigenesis may provide clues for detecting precancerous states [133]. Application of such approaches to iPSC-derived lung organoids or patient-specific lung cancer organoids offers a personalized framework, as these models originate from the same individual and may recapitulate patient-specific molecular features.

Concluding remarks

In vitro culture systems are rapidly evolving to more accurately model human pathophysiology, including cancer, thereby addressing many historical limitations. When integrated with advanced single-cell and multi-omics sequencing technologies and precise genome engineering tools, lung organoid models offer unprecedented opportunities to elucidate mechanisms of cell plasticity and lineage dynamics during lung cancer evolution. Insights derived from these systems may inform the development of interventional strategies aimed at suppressing oncogenic transformation before overt malignancy emerges. Ultimately, continued refinement of lung organoid platforms has the potential to expand therapeutic strategies and advance personalized prevention and treatment approaches for lung cancer.

NOTES

Conflict of interest

Jong-Il Kim has been an editor-in-chief of Organoid since 2023. However, Jong-Il Kim was not involved in the review process of this manuscript.

Funding

This research was supported by a grant from the MD-PhD/Medical Scientist Training Program through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea. This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MSIT) (2020R1A2C3012524). This research was supported by Basic Science Research Program through the NRF funded by the Ministry of Education (2020R1A6A1A03047972). This research was supported by a Korea Basic Science Institute (National Research Facilities and Equipment Center) grant funded by the Ministry of Education (2021R1A6C101A445). The manuscript includes revisions to parts of the first author’s PhD thesis, “Assessment of carcinogen exposure among cooks and modeling lung cancer initiation using human airway organoids” [100].

Authors’ contributions

Conceptualization: SM; Data curation: SM, DL; Funding acquisition: JIK; Methodology: SM; Project administration: JIK; Visualization: SM; Resources: DL, JC, SL; Supervision: JIK; Writing–original draft: SM, DL; Writing–review & editing: all authors.

Data availability

Not applicable.

Fig. 1.

Schematic of normal lung epithelial differentiation and plasticity in cancer evolution (A) and a lung cancer evolution model using lung organoids (B) (small cell lung cancer [SCLC]; A, ASCL1; N, NEUROD1; P, POU2F3; Y, YAP1; LUSC, lung squamous cell carcinoma; LUAD, lung adenocarcinoma). Illustrated by the authors and created with BioRender.

Table 1.

Genetically engineered lung organoid models of lung cancer evolution

Lineage	Study	Source	Engineered genes	Transformation^a)
Alveolar	Dost et al. [39]	iPSC (human)	KRAS^G12D [AAVS1-TetO–, rtTA]	Not reported
		ASC (mouse)	Kras^G12D [LSL–], Trp53^f/f	Tumor (LUAD)
	Chaudhary et al. [40]	ASC (mouse)	Kras^G12D [LSL–], Sftpc^CreER	Tumor (LUAD)
	Moye et al. [41]	ASC (mouse)	Kras^G12D [LSL–], Trp53^f/f	Tumor (LUAD)
	Han et al. [42]	ASC (mouse)	Gprc5a^–/–, Sftpc^CreER	Not reported
	Miura et al. [43]	iPSC (human)	HER2 [AAVS1-TetO-rtTA–]	Not reported
	Ebisudani et al. [44]	ASC (human)	TP53, CDKN2A [sgRNA], EGFR^L858R, KRAS^G12D [OV]	Not reported
	Hill et al. [45]	ASC (mouse)	EGFR^L858R [TetO–], rtTA [LSL–]	Not reported
	Naranjo et al. [46]	ASC (mouse)	Trp53^f/f, Kras^G12D[LSL–]	Tumor (LUAD)
			Trp53 [sgRNA], Braf^V600E [LSL–]
			Trp53^f/f, Eml4/Alk [sgRNAs]
	Gardner et al. [47]	ASC (mouse)	Myc^T58A [H11-LSL–], Sftpc^Cre-ERT2	No tumor
	Chen et al. [48]	ASC (mouse)	Egfr^{T790M, L858R}[Col1A1-LSL–]	Tumor (LUAD)
Airway	Gardner et al. [47]	ASC (mouse)	Myc^T58A [H11-LSL–], Rb1^f/f, Trp53^f/f, Ascl1^Cre–ERT2;	Tumor (SCLC)
Airway	Gardner et al. [47]	ASC (mouse)	Myc^{WT, T58A}[TetO–], rtTA3 [R26-LSL–], Ascl1^Cre-ERT2	Tumor (SCLC)
	Kim et al. [49]	ASC (human)	SOX2 [OV]	No tumor^b)
	Jeong et al. [50]	ASC (mouse)	Trp53^f/f, Keap1^f/f, Krt5^Cre-ERT2	Not reported
	Lazarus et al. [51]	ASC (mouse)	Bcl11a [LSL–]; Bcl11a^f/f, Cre–ERT2 [R26–]	Not reported
	Liu et al. [52]	ASC (mouse)	Trp53^f/f, SOX2, PRKCI, ECT2 [OV]	Tumor (LUSC)
	Hai et al. [53]	ASC (mouse)	sgRNA^{Pten, Trp53, CDKN2A}, Sox2 [R26-LSL–]	Tumor (LUSC)
	Pan et al. [54]	ASC (mouse)	Trp53^f/f, Kmt2d [sgRNA]	Tumor (LUSC)
			Trp53^f/f, Pten [sgRNA]	Tumor (LUAD)
	Park et al. [55]	ASC (human)	RB1 [shRNA], TP53[DN], MYC, BCL2, myrAKT [OV]	Tumor (SCLC)
	Chen et al. [48]	ASC (mouse)	Egfr^{T790M or L858R} [Col1A1-LSL–]	No tumor
Both	Chen et al. [48]	ASC (mouse)	Egfr^{T790M or L858R} [Col1A1-LSL–]	Tumor (LUAD+LUSC)
Cancer	Tong et al. [56]	LUAD (mouse)	Kras^G12D [LSL–], Lkb1^f/f	Tumor (LUSC)
	Qin et al. [57]	LUAD (mouse)	EML4-ALK^{L1196M, WT} [LSL–]	Tumor (LUSC)
	Ebisudani et al. [44]	LUAD (human)^c)	sgRNA^NKX2-1; NKX2-1 [TetO–, rtTA]	Tumor (LUAD)

iPSC, induced pluripotent stem cell; ASC, adult stem cell; CreER, Cre recombinase fused with estrogen receptor; TetO, tetracycline operator; rtTA, reverse tetracycline transactivator; AT2, alveolar type 2; AT1, alveolar type 1; LSL, LoxP-stop-LoxP cassette; LUAD, lung adenocarcinoma; sgRNA, single-guide RNA; OV, overexpression; KO, knockout; SCLC, small cell lung cancer; R26, Rosa26; LUSC, lung squamous cell carcinoma; shRNA, short hairpin RNA; DN, dominant negative form; myr, myristoylated.

^a)Demonstrated by a transplantation experiment in mice.

^b)Showed squamous metaplasia.

^c)Patient-derived organoids.

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