Cancer cells extensively reprogram their metabolism to ensure survival, proliferation, and fulfillment of biosynthetic demands. In this context, three major metabolic paradigms emerge: glutaminolysis, the Warburg effect, and the lactate shuttle. However, these canonical models remain insufficient to fully explain certain critical aspects of tumor metabolism, particularly the direction of carbon flux in normoxic tumor regions, compartment-specific redox regulation, and nutrient-driven mechanisms that support epigenetic programs.

Glutamine is an essential metabolite for rapidly dividing cells and contributes to nucleotide, amino acid, and lipid biosynthesis (1). Its uptake is enhanced through Solute Carrier Family 1 Member 5 (SLC1A5, ASCT2) and the sodium-dependent neutral amino acid transporters SNAT1 and SNAT2 (2). Once inside the cell, glutamine is converted to glutamate by glutaminase (GLS) and subsequently transformed into α-ketoglutarate (α-KG) via glutamate dehydrogenase (GLUD1) or aminotransferases. α-KG then enters the tricarboxylic acid (TCA) cycle, supporting both energy production and anabolic metabolism (3). Moreover, the MYC oncogene enhances GLS1 expression, thereby promoting glutamine dependence (4). GLUD1 also contributes to NADPH production, helping maintain redox homeostasis under oxidative stress conditions (5).

Glutamine additionally serves as a nitrogen donor in purine and pyrimidine biosynthesis. Thus, glutaminolysis represents a central pathway supporting energy metabolism, biosynthetic precursor generation, and oxidative defense mechanisms (6). Recent studies have shown that cancer cells exhibit metabolic flexibility by utilizing diverse energy sources such as glutaminolysis and β-oxidation (7).

The Warburg effect refers to the conversion of glucose to lactate in the cytosol despite the presence of sufficient oxygen and functional mitochondria, rather than its complete oxidation to acetyl-CoA. This metabolic shift provides three key advantages:

Lactate and protons produced by hypoxic and normoxic tumor cells are transported bidirectionally across the plasma membrane by monocarboxylate transporters (MCTs), depending on the concentration gradient. Among the SLC16 gene family, MCT1 (SLC16A1), MCT2, MCT3, and MCT4 isoforms are particularly relevant (13, 14).

In hypoxic tumor cells, elevated glycolytic flux markedly increases lactate production. Under these conditions, hypoxia-inducible factor-1 alpha (HIF-1α) becomes activated and induces MCT4 expression. Although MCT4 exhibits low affinity for lactate (Km ≈ 20–35 mM), its high transport capacity allows efficient efflux of lactate and protons into the ECM. This process prevents intracellular acidosis and ensures glycolytic continuity (13, 15).

While a portion of extracellular lactate enters systemic circulation, another fraction is re-imported into normoxic cancer cells via upregulated MCT1 (Km ≈ 3–5 mM) (14, 16).

In normoxic cancer cells, enhanced glycolysis, increased lactate shuttle activity, and inhibition of the pyruvate dehydrogenase (PDH) complex collectively elevate cytoplasmic lactate levels. Intracellular lactate accumulation reverses the direction of the lactate dehydrogenase (LDH) reaction, shifting flux from pyruvate → lactate toward lactate → pyruvate. During this process, lactate is converted into pyruvate, accompanied by the reduction of NAD⁺ to NADH (17).

Cytosolic NADH cannot directly cross the mitochondrial inner membrane. Therefore, it transfers its reduced electrons into mitochondria through the malate–aspartate shuttle (MAS) and glycerol-3-phosphate shuttle (G3P) systems. Electrons carried via MAS are transferred to mitochondrial NAD⁺, while the G3P shuttle mediates electron transfer through FAD. In this way, electrons derived from NADH and FADH₂ are delivered to the electron transport chain and contribute to ATP synthesis via oxidative phosphorylation (18). These mechanisms serve to maintain cytosolic NAD⁺/NADH homeostasis.

In both shuttle systems, the regenerated cytosolic NAD⁺ supports numerous NAD⁺-dependent anabolic processes such as glycolysis, serine/glycine biosynthesis, folate metabolism, lipogenesis, and nucleotide synthesis (19). MAS is superior in terms of energy efficiency and is predominantly used in normoxic cancer cells. In contrast, the G3P shuttle functions as an alternative redox mechanism particularly in highly proliferative tumor cells. Recent literature reports that the components of the G3P shuttle are overexpressed especially in hepatocellular carcinoma, glioblastoma, and prostate cancer, and that this condition is associated with tumor progression and metabolic flexibility (20).

In normoxic cancer cells, pyruvate arises from several sources, particularly accelerated glycolysis and the lactate shuttle. The mitochondrial pyruvate dehydrogenase complex (PDH), however, is inhibited by phosphorylation through HIF-1–induced pyruvate dehydrogenase kinases (PDKs). This inhibition restricts the conversion of pyruvate into acetyl-CoA and reduces oxidative flux, thereby contributing to cytoplasmic pyruvate accumulation (9). In a single cell, glycolysis and gluconeogenesis rarely function at full capacity at the same time. In tumors, however, spatiotemporal heterogeneity enables certain steps of gluconeogenesis, especially mitochondrial PEPCK/PCK2, to remain active. This “partial gluconeogenesis” does not culminate in glucose production. Instead, it provides biosynthetic intermediates such as phosphohydroxy acids, ribose-5-phosphate, and the glycerol backbone (21). Therefore, in cancer, gluconeogenesis is more accurately interpreted as “flux/intermediate supply–oriented” rather than “product–oriented” (22).

The principal alternative pathway for pyruvate entry into mitochondria is its anaplerotic conversion to oxaloacetate (OAA) via pyruvate carboxylase (PC); the primary role of PC is to expand the TCA intermediate pool and feed anabolic requirements such as lipogenesis through citrate export, and aspartate/thymidylate synthesis (23, 24). Clinical and experimental data demonstrate that PC expression/activity is increased in NSCLC, breast, and certain other cancers, and that PC suppression reduces proliferation, colony formation, and tumor burden (24, 25).

Alterations in glutaminolysis and fatty acid β-oxidation dynamically influence the requirement for PC. An increase in β-oxidation, accompanied by a rise in acetyl-CoA and NADH, further suppresses PDH and redirects pyruvate flux toward PC-mediated anaplerosis. In some models, glutaminolysis cannot fully compensate for the loss of PC, and dependency on PC for anaplerosis persists (26). In this context, PCK2-mediated “partial gluconeogenesis” and PC-mediated anaplerosis emerge as complementary strategies acting together not to synthesize glucose but to provide redox balance and biosynthetic precursors (27).

Mitochondrial oxaloacetate (OAA) cannot be directly transported out of mitochondria; therefore, it is converted into different metabolites such as citrate, malate, aspartate, or phosphoenolpyruvate (PEP), which are then transferred to the cytoplasm via specific carrier proteins.

One of the most important metabolic pathways involving mitochondrial oxaloacetate (OAA) is citrate synthesis. OAA condenses with acetyl-CoA via citrate synthase (CS). The product of this reaction is citrate. Depending on the immediate requirements of the cell, citrate may continue oxidative flux in the TCA cycle. The excess is exported from the mitochondria into the cytosol via SLC25A1—citrate carrier (CIC) located in the inner mitochondrial membrane (28, 29). In cancer, expression and/or activity of CS is frequently increased (e.g., in ovarian and prostate cancer) (30). SLC25A1/CIC levels are also elevated in many tumors, suggesting that citrate efflux from mitochondria is increased. High SLC25A1 has been associated with poor prognosis in pan-cancer analyses (31). Moreover, SLC25A1 is a transcriptional target of mutant p53 (32).

Another major fate of mitochondrial oxaloacetate (OAA) is conversion to malate. This reaction is reversibly catalyzed by malate dehydrogenase-2 (MDH2) in the mitochondrial matrix and influences TCA flux. During the reduction of OAA to malate, NADH is oxidized to NAD⁺, thereby increasing the mitochondrial NAD⁺/NADH ratio (33). The resulting malate may continue in the TCA cycle (34). The excess is exported to the cytosol via SLC25A11/OGC (malate ⇆ 2-oxoglutarate) or SLC25A10/DIC (malate ⇆ phosphate) carriers (35, 36). This axis is reprogrammed in cancer. In prostate cancer, MDH2 is elevated and associated with docetaxel resistance. In contrast, MDH2 is decreased in clear-cell renal carcinoma and affects sensitivity/ferroptosis response (37). SLC25A10 is overexpressed in many models; its suppression weakens proliferation and tumorigenesis. Germline loss-of-function variants of SLC25A11 confer hereditary predisposition to paraganglioma/pheochromocytoma (38, 39).

Another important metabolic fate of mitochondrial oxaloacetate (OAA) is its conversion to aspartate via Glutamate Oxaloacetate Transaminase 2 (GOT2). The resulting aspartate is exported from the mitochondria to the cytosol by the inner membrane carriers SLC25A12 (AGC1) and SLC25A13 (AGC2/citrin); this step is part of the malate–aspartate shuttle (MAS) and enables carbon/nitrogen flow between compartments (40).

Export of aspartate through AGC1/AGC2 allows the transfer of cytosolic→matrix NADH equivalents via MAS; under ETC inhibition/hypoxia, decreased aspartate limits proliferation (41). In addition, aspartate efflux serves cataplerotic utilization of TCA intermediates and contributes to balancing tissue-specific carbon/nitrogen fluxes (34). In the cancer context, this pathway is frequently upregulated; GOT2 and Aspartate–Glutamate Carriers (AGC1/AGC2) may enhance aspartate supply to support growth, whereas inhibition of GOT2 or blockade of aspartate transport can limit cell proliferation and tumor growth (depending on model and context) (40, 42).

Under certain cellular stress conditions or proliferative signals, mitochondrial oxaloacetate (OAA) can also be converted into phosphoenolpyruvate (PEP) and directed into the cytosol. This conversion is catalyzed by mitochondrial phosphoenolpyruvate carboxykinase-M (PEPCK-M; PCK2) in the mitochondrial matrix. During the reaction, OAA undergoes decarboxylation and phosphorylation using GTP, generating PEP and GDP (43). The resulting GDP molecules contribute to maintaining the mitochondrial GDP/GTP balance and also serve as a required cofactor for succinyl-CoA synthetase, one of the key enzymes of the TCA cycle, thereby supporting cycle continuity (44).

PEPCK-M (PCK2) expression has been shown to be significantly increased in pancreatic, lung, and colorectal tumors (43). This increase provides metabolic flexibility to cancer cells by supporting carbon flux and energy production independent of glucose. On the other hand, pharmacological or genetic inhibition of PCK2 disrupts this adaptation, reducing cell proliferation and increasing metabolic stress (45).

The identity of the carrier responsible for transporting PEP into the cytosol has not yet been fully clarified; however, recent studies suggest that this transfer occurs largely through SLC25A1 and possibly with indirect contributions from other mitochondrial carriers (46, 47). PEP reaching the cytoplasm is used not for gluconeogenesis but as a substrate for anabolic and redox-balancing pathways such as serine/glycine biosynthesis, purine/pyrimidine production, and NADPH synthesis (43, 45).

It has been shown that in lung adenocarcinomas and tumors with growth capacity under glucose-deprived conditions, mitochondrial export of PEP is accelerated, thereby directing OAA toward non-gluconeogenetic anabolic pathways (46). In this respect, cytosolic anabolic utilization of PEP derived from OAA is considered an important metabolic reprogramming strategy that reduces glucose dependence of cancer cells and supports tumor progression (43). This allows cancer cells to enhance NADPH production, redox homeostasis, and biosynthetic capacity by exploiting alternative carbon sources.

Citrate transported into the cytosol from mitochondria via the SLC25A1 carrier is cleaved by ATP-citrate lyase (ACLY) into acetyl-CoA and oxaloacetate (47). This reaction is ATP-consuming and represents an important anabolic switch upregulated in cancer (48). ACLY is highly expressed particularly in cells activated by oncogenic pathways such as PI3K/Akt/mTOR and KRAS, and this increase facilitates not only lipid synthesis but also epigenetic reprogramming processes (49).

Malate exported to the cytosol is reconverted into oxaloacetate (OAA) and NADH by cytosolic malate dehydrogenase (MDH1) using NAD⁺. The resulting cytosolic OAA is converted to phosphoenolpyruvate (PEP) by PEPCK-C and enters numerous metabolic pathways.

In cancer: Expression and activity of MDH1 are frequently elevated. For example, in pancreatic and lung cancers, overexpression of MDH1 facilitates the conversion of malate to OAA, thereby enhancing PEP production and anabolic metabolism (62). This increase not only directs carbon flux toward serine/glycine biosynthesis and nucleotide synthesis via PEP but also supports cytosolic redox homeostasis through NADH production. NADH can be directed toward lactate production via lactate dehydrogenase (LDH), maintaining the NAD⁺/NADH balance and contributing to the energy requirements of tumor cells with high glycolytic rates (37). Excessive MDH1 activity has also been shown to increase proliferation in cancer and strengthen cellular resistance to metabolic stress (63). Therefore, MDH1 is considered a nodal point with both biosynthetic and redox-regulatory functions in certain tumor types.

Malate transported into the cytosol is converted by the NADP⁺-dependent malic enzyme (ME1) through oxidative decarboxylation into pyruvate + CO₂ + NADPH; this reaction directly feeds the cytosolic NADPH pool (64).

NADPH produced by ME1 is used by Fatty Acid Synthase (FASN) in fatty acid synthesis and as an essential electron donor in antioxidant defense via the glutathione and thioredoxin systems; thus, ME1 flux supports both lipogenesis and oxidative stress tolerance (65).

This flux generally follows the pathway citrate → ACLY → OAA → (MDH1) → malate → (ME1) → pyruvate + NADPH; pyruvate can then re-enter the mitochondria to be directed toward acetyl-CoA by PDH or OAA by PC, thereby ensuring integration between cytoplasmic and mitochondrial carbon and reducing equivalents (47).

In cancer, particularly in KRAS-dependent pancreatic adenocarcinoma, the Glutamate Oxaloacetate Transaminase 1 (GOT1)-MDH1-ME1 axis is reprogrammed to enhance NADPH production; silencing GOT1 or suppressing ME1 leads to intracellular ROS accumulation, redox imbalance, and loss of proliferation (66).

NADPH supplied by ME1, together with NADPH derived from the pentose phosphate pathway (G6PD) and Isocitrate Dehydrogenase 1 (IDH1), forms complementary reservoirs for lipid biosynthesis and redox homeostasis in tumors; their relative contributions vary depending on cell type and oncogenic drivers (65).

Aspartate transported into the cytosol via AGC1/2 carriers participates in various metabolic pathways. The reconversion of aspartate into OAA in the cytosol is catalyzed by AST/GOT1; this process has been discussed in detail above under the section on OAA metabolism (55). Beyond this, aspartate is a critical metabolite particularly for nucleotide biosynthesis and nitrogen metabolism.

Aspartate is an essential substrate for both purine and pyrimidine biosynthesis. In purine biosynthesis, aspartate contributes as a nitrogen donor to the purine ring during the later stages of inosine monophosphate (IMP) synthesis. In pyrimidine synthesis, it combines with carbamoyl phosphate to initiate the orotate pathway, contributing to uridine monophosphate (UMP) formation (61). Therefore, in proliferative cells, aspartate availability is considered a limiting factor for DNA/RNA synthesis. Indeed, under conditions of electron transport chain inhibition, mitochondrial aspartate production decreases, cytosolic aspartate levels fall, and tumor cell proliferation ceases (41).

Another important fate of aspartate is its involvement in the urea cycle. In the cytosol, argininosuccinate synthase (ASS1) combines aspartate with citrulline to form argininosuccinate. This step is critical for the detoxification of nitrogen as urea in cancer cells with accelerated amino acid catabolism. However, in many tumors, ASS1 expression is suppressed. This results in the diversion of aspartate away from the urea cycle and toward nucleotide biosynthesis, thereby enhancing the proliferative capacity of the tumor (67). Thus, aspartate is under dual metabolic pressure: tolerating the nitrogen load imposed by increased amino acid catabolism on the one hand, and sustaining DNA/RNA synthesis on the other.

In conclusion, aspartate exported into the cytosol is not limited to conversion into OAA. Through nucleotide biosynthesis, the urea cycle, and the malate–aspartate shuttle, it provides both the building blocks necessary for cell division and contributes to the maintenance of nitrogen and redox balance. In cancer, accelerated amino acid catabolism further increases aspartate demand by accelerating urea metabolism. Therefore, aspartate production and transport represent a limiting factor for proliferation and a central node in the reprogramming of cancer metabolism (66).

The Cancer-Induced Lactate Load and Oncologic Remodeling (CILLO) Hypothesis offers a new and original perspective on cancer metabolism. According to this hypothesis, the fate of pyruvate is not limited solely to its reduction to lactate. Lactate is not merely an energy substrate, an immune-evasion tool, or a waste product; rather, it functions as a critical metabolite in biosynthetic and redox processes, as well as in mechanisms that shape tumor prognosis.

The Warburg effect, glutaminolysis, and the lactate shuttle have largely been addressed separately in the literature. The CILLO Hypothesis not only integrates these three axes within a common framework. It also provides an expanded perspective that includes additional metabolic orientations such as pyruvate carboxylase–based anaplerosis, NADPH-producing pathways, and the serine/glycine–folate cycle. Furthermore, it incorporates epigenetic acetylation and the biosynthesis of phospholipids, fatty acids, cholesterol, and isoprenoids via citrate–acetyl-CoA. In this way, it proposes a comprehensive model that explains the metabolic flexibility unique to cancer cells.

In hypoxic cells, the reduction of glucose to lactate ensures NAD⁺ regeneration. In normoxic cells, lactate is converted to pyruvate and then to oxaloacetate (OAA) via pyruvate carboxylase. OAA is directed to different biosynthetic and redox pathways:

The CILLO Hypothesis presents a new paradigm that goes beyond long-described metabolic processes in the field of cancer biology by uniting these mechanisms within a holistic network framework. Although pathways such as the Warburg effect, glutaminolysis, and the lactate shuttle have been studied in detail for decades, with comprehensive molecular mechanisms proposed for each, these studies have mostly focused on individual metabolites, specific cell types, or isolated pathways. Thus, although the literature is rich, it is fragmented and dispersed.

The contribution of the CILLO Hypothesis is to integrate this fragmented information at the systems level. It interprets the metabolic orientations of cancer cells along the lactate–pyruvate–OAA axis not only in terms of energy production, but also within the broader context of anabolic growth, redox homeostasis, nucleotide synthesis, and microenvironmental adaptation.

We can gather the original contributions of this hypothesis to the literature under the following headings:

Different organ tumors utilize lactate metabolism in unique contexts. In glioblastoma, lactate contributes to immune evasion by suppressing CD8⁺ T-cell functions and by creating an immunosuppressive microenvironment through histone lactylation. In the liver, “glucose-non-producing gluconeogenesis” via PCK2 supplies carbon to serine/glycine and nucleotide biosynthesis and has been associated with proliferation and poor prognosis in hepatocellular carcinoma. In lung and breast cancers, overexpression of PC is decisive in meeting the needs for lipogenesis and anaplerosis by directing pyruvate to OAA. These organ-specific variations indicate that the CILLO hypothesis may have heterogeneous clinical implications across different tumor types.

The CILLO Hypothesis opens new horizons in the development of therapeutic strategies by revealing that the metabolic orientations executed by cancer cells along the lactate, pyruvate, and OAA axis are not only biological but also clinically targetable processes. The holistic metabolic model offered by this hypothesis provides a foundation for going beyond current treatments to break cancer’s metabolic flexibility, limit tumor adaptation, and target specific biosynthetic needs.

Combining metabolic targets with immunotherapy and classical chemotherapies increases the clinical relevance of the CILLO axis. For example, MCT1 inhibition with AZD3965, when considered together with programmed cell death protein 1 blockade, may reduce immune suppression; PC and PCK2 inhibition may show synergy with antifolate therapies; and BPGM modulation may regulate erythrocyte oxygen delivery and disrupt hypoxia adaptation. These combination strategies offer a new framework for personalized treatments following tumor type–specific metabolic profiling.

By proposing that cancer cells use lactate, pyruvate, and OAA metabolism as an integrated network, the CILLO Hypothesis enables the evaluation of metabolites and associated enzymes in this axis as biomarkers. In particular, the fact that these molecules are directly related to the tumor cell’s capacity for metabolic adaptation suggests that biochemical indicators based on the CILLO hypothesis may provide important clinical signals for diagnosis, prognosis, and treatment response. Findings that appear in a fragmented fashion in the literature can be placed into a systematic framework with this hypothesis, allowing the potential biomarkers to be evaluated in a logical context. In this scope, possible biomarkers that may be observed in cancerous tissue or in circulating biological fluids include plasma lactate/pyruvate ratio, MCT1/MCT4 expression ratio, pyruvate carboxylase (PC) activity, glycerol-3-phosphate dehydrogenase (GPD1) expression, and citrate/malate/aspartate levels. Each of these markers can provide information about the tumor cell’s substrate utilization, mitochondrial/cytosolic fluidity, oxygenation status, and biosynthetic orientation. Furthermore, when these parameters are made monitorable by non-invasive methods such as liquid biopsy, they may become valuable tools for early diagnosis in cancer, monitoring of treatment response, or evaluating the efficacy of metabolically targeted therapies.

The clinical validity of biomarkers based on the CILLO hypothesis has been demonstrated in studies conducted in recent years. In glioblastoma, the lactate–immune signature shows potential for predicting response to immunotherapy. In hepatocellular carcinoma, PCK2 levels have been associated with proliferation and prognosis. In breast and lung cancers, PC expression correlates with tumor burden and treatment resistance. These findings indicate that biomarkers within the CILLO axis may have tumor type–specific diagnostic and prognostic value.

Multilayered experimental approaches are required to validate the metabolic orientations and anabolic/redox contributions along the lactate–pyruvate–OAA axis proposed by the CILLO Hypothesis. In this regard, in cell culture models under different oxygen concentrations and substrate limitations, the metabolic flexibility of cancer cells can be tested. Tracking metabolites such as glucose, glutamine, and lactate with labeled isotopes (e.g., ¹³C or ¹⁵N) enables quantitative monitoring of carbon flux directions through LC–MS/MS–based stable isotope tracing (SIRM) analyses.

At the same time, expression levels and activities of critical enzymes such as ATP-citrate lyase (ACLY), pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), GAPDH, ME1, LDHB, MCT1, and SLC25A1 can be analyzed by qRT-PCR (gene level), Western blot and ELISA (protein level), and immunoprecipitation assays. Enzyme activity measurements can be performed with colorimetric or fluorometric spectrophotometric assays that track NADH/NAD⁺ or NADP⁺/NADPH production.

In addition, outputs such as intracellular redox status (NADPH/NADP⁺, GSH/GSSG ratio), cell division rate, mitochondrial function, lipid synthesis, and epigenetic modifications (e.g., histone acetylation, lactylation) can also be measured quantitatively under cell culture conditions, enabling the hypothesis to be tested at the systems level.

The CILLO Hypothesis posits that cancer cell metabolism is not merely a system focused on energy production, but rather an adaptable and versatile network that is highly responsive to environmental conditions. This hypothesis reveals that the metabolic orientations shaped around the lactate–pyruvate–oxaloacetate (OAA) axis cannot be evaluated solely within the framework of the classical Warburg effect or glutaminolysis; instead, these metabolites play multilayered roles in epigenetic regulation, redox balance, and anabolic synthesis pathways.

In the current literature, it is widely accepted that lactate is active not only in energy metabolism but also in processes such as immune suppression, reprogramming of stromal cells, and epigenetic modifications (e.g., histone lactylation). Similarly, overexpression of pyruvate carboxylase in certain cancer types and the production of OAA have been shown to mediate gluconeogenesis-like anabolic fluxes by directing NAD⁺ regeneration, aspartate biosynthesis, and serine/glycine/phospholipid metabolism (68). By addressing these metabolic orientations not in a fragmented manner but through a holistic approach, the CILLO Hypothesis explains the reprogramming of the cancer cell at the systems level.

The originality of the hypothesis lies in proposing that lactate contributes to energy production by being transported from hypoxic to normoxic regions. In addition, in normoxic cancer cells, lactate participates in gluconeogenesis-like reactions through the production of mitochondrial OAA and phosphoenolpyruvate (PEP). These processes support the generation of intermediate metabolites. These orientations both ensure the sustainability of glycolytic flux by providing NAD⁺ regeneration and maintain redox homeostasis to establish a biochemical substrate favorable for proliferation.

The CILLO Hypothesis also reveals that this metabolic network offers potential therapeutic targets (e.g., MCT1, PC, GAPDH, ME1) and biomarker candidates (e.g., cytosolic PEP, OAA, NADPH/NADP⁺ ratio). Thus, it may contribute not only to a better understanding of cancer but also to the development of personalized treatment approaches.

In conclusion, the CILLO Hypothesis offers a new paradigm based on the lactate–pyruvate–OAA axis that integrates mechanisms dispersed in the existing metabolic literature. This holistic approach shows that cancer metabolism is not solely focused on energy production but is also reorganized to serve biosynthetic and redox needs. The hypothesis provides an important framework that can lead to new research in both basic science and translational oncology.

This figure depicts the integrated metabolic adaptations of a normoxic cancer cell, highlighting the central role of lactate in biosynthetic pathways through the CILLO Hypothesis (). Lactate imported via MCT1 () is converted to pyruvate by LDHB. Instead of fueling oxidative metabolism through PDH, pyruvate is redirected to oxaloacetate (OAA) by mitochondrial pyruvate carboxylase (PC). OAA and its derivatives support multiple anabolic processes: malate can generate cytosolic NADPH via malic enzyme (ME1), citrate export and ATP-citrate lyase (ACLY) fuel lipid and cholesterol synthesis, and phosphoenolpyruvate (PEP) links to serine/glycine and folate metabolism as well as aspartate-derived nucleotide synthesis.

Meanwhile, glucose metabolism proceeds through glycolysis and the Warburg pathway (), with diversion into the pentose phosphate pathway () to generate ribose-5-phosphate and NADPH for nucleotide biosynthesis and redox balance. In parallel, glutaminolysis () sustains TCA cycle anaplerosis via α-ketoglutarate, ensuring metabolic flexibility.