It was shown that the content of individual amino acids changes in breast cancer by an average of 10–15% compared with healthy controls. For some amino acids (Thr, Arg, Met, and Ser), an increase in concentration is more often observed in breast cancer, and for others, a decrease is observed (Asp, Pro, Trp, and His). The accuracy of diagnostics using individual amino acids is low and increases when a number of amino acids are combined with each other or with other metabolites. Gln/Glu, Asp, Arg, Leu/Ile, Lys, and Orn have the greatest significance in assessing the risk of breast cancer.
In different studies, both the contents of individual amino acids and the complete amino acid profile of plasma and/or serum were evaluated. It has been shown, for example, that the arginine levels were significantly lower and the ornithine levels were significantly higher in breast cancer patients than in control patients [44][64]. Differences between breast cancer cases and controls were observed for the levels of Trp, Thr, Ala, Gly, Pro, Ile, Leu, and Val [34]. Miyagi et al. observed significant changes in the amino acid profiles between cancer patients and controls. They found that the concentrations of Thr, Pro, Ser, Gly, Ala, and Orn were significantly higher, but the concentrations of Gln, Trp, His, Phe, and Tyr were significantly lower compared to the control. Compared with the data from Lai et al., they found similarities in decreasing His and Gln levels and increasing Pro and Ala levels in breast cancer patients. Budczies et al. demonstrated that changes in the amino acid metabolism were associated with at least a 1.9-fold increase in 16 amino acids in breast cancer compared with healthy breast tissue [45][46][50,65].
To compare the change in the contents of individual amino acids in breast cancer according to different authors, scholars compared the relative contents of amino acids in relation to healthy controls. It can be seen that a significant change in the concentration is rarely observed, for example, an increase in the contents of Arg [47][48][52,56], Thr [47][52], and Ser [33] by 2 times, a decrease in the content of Asp by 1.5 times [48][49][46,56], etc. In most cases, the change in concentration is up to 10–15% compared to the control.Some essential amino acids (EAAs)—Trp, His, Met, and branched-chain amino acids—are directly associated with tumor growth and treatment resistance [112][113][114][115][180,181,182,183]. EAAs provide important raw materials for protein synthesis, influence the biological behavior of cells, and induce therapeutic resistance in breast cancer cells. Strekalova et al. [116][184] found that Met plays a critical role in maintaining the self-renewal and survival of cancer stem cells. Met restriction reduced the population of cancer stem cells in TNBC. Saito et al. [117][185] showed that Leu not only promotes cell proliferation in ER-positive breast cancer, but also participates in the mechanism of resistance to tamoxifen. In addition, essential amino acids, as important nutrients, mediate the development of an immunosuppressive tumor microenvironment in breast cancer. For example, kynurenine, a product of Trp metabolism, can inhibit T cell proliferation and differentiation, leading to immune evasion and tumor progression in breast cancer [118][186]. Interestingly, both Trp and kynurenine were lower in plasma in breast cancer patients compared to controls, especially in women with estrogen-receptor-negative and advanced breast cancers [119][187]. These results show an intrinsic relationship between EAA metabolism and the immune microenvironment in breast cancer.
Under physiological conditions, Gln is transported into cells by many transporters such as Ala, Ser, Cys-preferential transporter 2 (ASCT2, also known as SLC1A5), and L-type amino acid transporter 1 (LAT1, also known as SLC7A5). In TNBC, both ASCT2 and LAT1 are overexpressed [83][120][122,190]. Compared to other subtypes of breast cancer, TNBC is more Gln dependent and sensitive to glutaminolysis-targeted therapy due to glutaminase overexpression (GLS) [121][122][191,192], which is associated with high-grade metastatic breast cancer. Several small-molecule GLS inhibitors, such as CB-839, have been developed to combat the dysregulation of glutaminolysis [123][193]. Other combination therapies, such as the combination of GLS inhibition and bevacizumab (an anti-angiogenesis monoclonal antibody targeting VEGF), also show antitumor effects on TNBC [124][194]. Ginsenoside was described to effectively inhibit TNBC by suppressing Gln uptake and Glu production by down-regulating glutaminase 1 (GLS1) expression [125][195]. Ginsenoside treatment further reduced cellular ATP production, decreased amino acid utilization associated with Gln metabolism, and induced glutathione depletion and reactive oxygen species accumulation, which consequently triggered apoptosis in TNBC.
Increased Glu production by GLS may support the uptake of exogenous cystine via the cystine/glutamate antiporter xCT to maintain redox balance. As a clinically approved anti-inflammatory drug, sulfasalazine (SASP) was found to inhibit xCT activity and retard the growth of TNBC [126][197]. By immunizing mice with a DNA-based vaccine expressing the xCT protein, the cell surface immunotargeting of the xCT antigen effectively attenuated tumor growth and lung metastasis, and increased chemosensitivity to doxorubicin [127][198].
In addition to Gln and Cys, TNBC cells are also somewhat dependent on the availability of several other amino acids such as Met, Asp, and Arg, suggesting that restricting these amino acids may have a therapeutic effect [128][200]. The depletion of either Met or Gln can increase the cell surface expression of the pro-apoptotic TNF-related apoptosis-inducing ligand receptor-2 (TRAIL-R2) and increase the sensitivity of TNBC cells to TRAIL-induced apoptosis [129][130][130,201]. Met deprivation in the diet increases cellular susceptibility to lexatumumab, an agonistic monoclonal antibody targeting TRAIL-R2, and reduces the rate of lung metastasis [131][202]. In addition, many tumor and stem cells depend on the biosynthesis of the universal methyl donor S-adenosylmethionine from the exogenous Met via methionine adenosyltransferase 2α (MAT2A) to maintain their epigenome [132][133][203,204]. A restriction on Met is enough to undermine the ability of TNBC to initiate a tumor, which is in part due to the impaired formation of S-adenosylmethionine. The combination of methionine restriction and the MAT2A inhibitor cycloleucine has a synergistic antitumor effect [116][184]. Under normal physiological conditions, the serum levels of Asp are lower than in the mammary gland, making Asp bioavailability a key regulator of circulating tumor cells and the metastatic potential of breast cancer. The restriction of Asp intake by the suppression of asparagine synthetase, treatment with L-asparaginase, or the restriction of Asp intake in the diet inhibits breast cancer metastasis [130][201]. The depletion of Arg by recombinant human arginase (rhArg) leads to the apoptosis of TNBC cells via reactive oxygen species and induces adaptive autophagy, while blocking the flow of autophagy via autophagy-targeting drugs enhances rhArg cytotoxicity [134][205]. The deprivation of Arg by L-arginase impairs tumor growth, leading to cell death [135][206]. Much attention has been paid to epigenetic modifications caused by enzymes of protein arginine methyltransferases, in which methylate Arg make a great contribution to the process of breast carcinogenesis and tumor suppression [136][207] and are targets for many types of cancer [137][208].
Leu uptake is predominantly mediated by the L-type amino acid transporter (LAT) family, a group of four Na+-independent transporters (LAT1, SLC7A5; LAT2, SLC7A8; LAT3, SLC43A1; and LAT4, SLC43A2) with an affinity for branched and neutral amino acid transporters [129][130]. Glutamine transport is largely mediated by Ala, Ser, and Cys-preferential transporter 2 (ASCT2; SLC1A5).
Amino acid deprivation (AADT) is becoming a promising strategy for the development of new therapeutic agents against cancer [138][211]. The rapid growth of tumors leads to a decrease in the expression of certain enzymes, which leads to the auxotrophy of some specific amino acids. Amino acid depletion selectively inhibits tumor growth because normal cells can synthesize amino acids through their normal machinery. The enzymes used in AADT are primarily obtained from microbes due to their easy availability. Thus, the deprivation of Gln leads to a decrease in cell proliferation and cell death in breast cancer cell lines [139][212]. Endocrine therapy is the standard treatment for estrogen-receptor-positive (ER+) breast cancer, but 40% of women experience a recurrence of the disease during therapy. A general analysis of transcription in cells revealed a suppression of the neutral and basic amino acid transporter SLC6A14, which is regulated by the increased expression of miR-23b-3p, which leads to impaired amino acid metabolism [140][213]. This altered cellular amino acid metabolism is supported by autophagy activation and the increased import of acidic amino acids (Asp and Glu) mediated by the SLC1A2 transporter. Targeting these amino acid metabolic dependencies increases the sensitivity of cells to endocrine therapy. The use of various drugs based on platinum nanoparticles leads to a disruption of the amino acid metabolism, a disruption of tRNA aminoacylation, and protein synthesis [141][216]. Mitrevska et al. [142][217] analyzed the effect on amino acid metabolism in MDA-MB-231 cells upon treatment with cisplatin, PtNP-10, and PtNP-40, and revealed a marked contrast between the effects of cisplatin and PtNP. The results indicate a higher sensitivity of MDA-MB-231 cells to PtNP compared to cisplatin, since the increase in the number of amino acids was associated with the degree of insensitivity to various chemotherapeutic agents [142][217]. In general, combinations of anticancer drugs and amino acids can improve the intratumoral distribution of the active substance and increase its bioavailability. In particular, amino acid-based poly(ether urea ester) (AA-PEUU), as a nanocarrier for the systemic delivery of gamboginic acid, demonstrates the effectiveness of engineered AA-PEUU nanocarriers with custom structures and universal customization for the systemic delivery of therapeutics in the treatment of TNBC [143][218]. Plasma amino acid analysis can be used to monitor treatment progress. So, Minet-Quinard et al. [144][47] showed that the plasma levels of Ser and Glu returned to normal six months after the surgical removal of the tumor. Dunstan et al. [145][219] considered the changes in the amino acid homeostasis during radiation therapy. The urinary histidine and alanine levels were shown to be elevated prior to radiotherapy, while the Thr, Met, Ala, Ser, Asp, and Gln levels were higher after 5 weeks of radiotherapy. Many complications such as cachexia, anorexia, and fatigue occur in the treatment of problems associated with breast cancer, and many studies have considered the addition of amino acids with BCAAs [146][220] or a single amino acid or its derivative [147][221] to reduce the effects of treatment.